Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-151_S2_L002_R1_001_val_1.fq.gz to EPI-151_S2_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-151_S2_L002_R1_001_val_1.fq.gz (24505793 sequences in total) Writing a G -> A converted version of the input file EPI-151_S2_L002_R2_001_val_2.fq.gz to EPI-151_S2_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-151_S2_L002_R2_001_val_2.fq.gz (24505793 sequences in total) Input files are EPI-151_S2_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-151_S2_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-151_S2_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-151_S2_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:1928:2248_1:N:0:ATCACG/1 77 * 0 0 * * 0 0 TAGTAGAGGTTTATATGTATTTATGTTTGGTTAGAAAAGGTTTATATGTATTTATGTTTGGTTAGTAGAGGTTTATATGTATTTATGTTTGGTTA BBBBBFFBFFBBFBBBBFFF/B/FBBF>> Writing bisulfite mapping results to EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:2114:2329:9695_1:N:0:ATCACG PGA_scaffold9__45_contigs__length_38581958 2 Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far Processed 23000000 sequence pairs so far Processed 24000000 sequence pairs so far 24505793 reads; of these: 24505793 (100.00%) were paired; of these: 16362174 (66.77%) aligned concordantly 0 times 3586050 (14.63%) aligned concordantly exactly 1 time 4557569 (18.60%) aligned concordantly >1 times 33.23% overall alignment rate 24505793 reads; of these: 24505793 (100.00%) were paired; of these: 16471528 (67.21%) aligned concordantly 0 times 3553354 (14.50%) aligned concordantly exactly 1 time 4480911 (18.29%) aligned concordantly >1 times 32.79% overall alignment rate Processed 24505793 sequences in total Successfully deleted the temporary files EPI-151_S2_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-151_S2_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 24505793 Number of paired-end alignments with a unique best hit: 9915442 Mapping efficiency: 40.5% Sequence pairs with no alignments under any condition: 12555282 Sequence pairs did not map uniquely: 2035069 Sequence pairs which were discarded because genomic sequence could not be extracted: 1 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 4881878 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 5033563 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 303264962 Total methylated C's in CpG context: 12323211 Total methylated C's in CHG context: 1414630 Total methylated C's in CHH context: 3533701 Total methylated C's in Unknown context: 52135 Total unmethylated C's in CpG context: 31194293 Total unmethylated C's in CHG context: 65160946 Total unmethylated C's in CHH context: 189638181 Total unmethylated C's in Unknown context: 646387 C methylated in CpG context: 28.3% C methylated in CHG context: 2.1% C methylated in CHH context: 1.8% C methylated in unknown context (CN or CHN): 7.5% Bismark completed in 0d 1h 37m 30s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-152_S3_L002_R1_001_val_1.fq.gz to EPI-152_S3_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-152_S3_L002_R1_001_val_1.fq.gz (29855018 sequences in total) Writing a G -> A converted version of the input file EPI-152_S3_L002_R2_001_val_2.fq.gz to EPI-152_S3_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-152_S3_L002_R2_001_val_2.fq.gz (29855018 sequences in total) Input files are EPI-152_S3_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-152_S3_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-152_S3_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-152_S3_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:1155:2233_1:N:0:CGATGT/1 77 * 0 0 * * 0 0 AATGGNTATGAAAATGTAATAAAGGAAGAGATATGAAAATGTAATAAAGGAANATTTATGANAATGTATTGTGGGAAGTGATATGAAAATGTAATATG BBBBB#B>> Writing bisulfite mapping results to EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far Processed 23000000 sequence pairs so far Processed 24000000 sequence pairs so far Processed 25000000 sequence pairs so far Processed 26000000 sequence pairs so far Processed 27000000 sequence pairs so far Processed 28000000 sequence pairs so far Processed 29000000 sequence pairs so far 29855018 reads; of these: 29855018 (100.00%) were paired; of these: 20215536 (67.71%) aligned concordantly 0 times 4311215 (14.44%) aligned concordantly exactly 1 time 5328267 (17.85%) aligned concordantly >1 times 32.29% overall alignment rate 29855018 reads; of these: 29855018 (100.00%) were paired; of these: 20302194 (68.00%) aligned concordantly 0 times 4294798 (14.39%) aligned concordantly exactly 1 time 5258026 (17.61%) aligned concordantly >1 times 32.00% overall alignment rate Processed 29855018 sequences in total Successfully deleted the temporary files EPI-152_S3_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-152_S3_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 29855018 Number of paired-end alignments with a unique best hit: 11727816 Mapping efficiency: 39.3% Sequence pairs with no alignments under any condition: 15750735 Sequence pairs did not map uniquely: 2376467 Sequence pairs which were discarded because genomic sequence could not be extracted: 0 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 5811638 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 5916178 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 358780662 Total methylated C's in CpG context: 14625118 Total methylated C's in CHG context: 1782524 Total methylated C's in CHH context: 4106805 Total methylated C's in Unknown context: 73648 Total unmethylated C's in CpG context: 34864391 Total unmethylated C's in CHG context: 74934034 Total unmethylated C's in CHH context: 228467790 Total unmethylated C's in Unknown context: 784418 C methylated in CpG context: 29.6% C methylated in CHG context: 2.3% C methylated in CHH context: 1.8% C methylated in unknown context (CN or CHN): 8.6% Bismark completed in 0d 1h 56m 1s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-153_S4_L002_R1_001_val_1.fq.gz to EPI-153_S4_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-153_S4_L002_R1_001_val_1.fq.gz (26009397 sequences in total) Writing a G -> A converted version of the input file EPI-153_S4_L002_R2_001_val_2.fq.gz to EPI-153_S4_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-153_S4_L002_R2_001_val_2.fq.gz (26009397 sequences in total) Input files are EPI-153_S4_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-153_S4_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-153_S4_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-153_S4_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:1785:2231_1:N:0:TTAGGC/1 77 * 0 0 * * 0 0 TGTTGNGATATATAATTGAAAATTGTTGAAAGTGTTATTAAATATAATGTAATAATAAATGATAAAGGGATGTAATGTTGTATATTAATTATTAT BBBBB#BBFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFBFFFFFFFFFFFFFFFFFFFFFFFFFFF YT:Z:UP D00743:144:CAAWNANXX:2:2207:1785:2231_2:N:0:TTAGGC/2 141 * 0 0 * * 0 0 ATANNNNNNNNNNNNNNNNNNNNNNNNNNNNTNNNATTTATTATTACATTATATTTAATAACACTTTCAACAATTTTCAATTATATATCACAACA BBB############################B###BB<>> Writing bisulfite mapping results to EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:1115:15676:64681_1:N:0:TTAGGC PGA_scaffold8__63_contigs__length_61151155 61151067 Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far Processed 23000000 sequence pairs so far Processed 24000000 sequence pairs so far Processed 25000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:2314:20445:24921_1:N:0:TTAGGC PGA_scaffold11__79_contigs__length_51449921 1 Processed 26000000 sequence pairs so far 26009397 reads; of these: 26009397 (100.00%) were paired; of these: 17345929 (66.69%) aligned concordantly 0 times 3790523 (14.57%) aligned concordantly exactly 1 time 4872945 (18.74%) aligned concordantly >1 times 33.31% overall alignment rate 26009397 reads; of these: 26009397 (100.00%) were paired; of these: 17425568 (67.00%) aligned concordantly 0 times 3786588 (14.56%) aligned concordantly exactly 1 time 4797241 (18.44%) aligned concordantly >1 times 33.00% overall alignment rate Processed 26009397 sequences in total Successfully deleted the temporary files EPI-153_S4_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-153_S4_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 26009397 Number of paired-end alignments with a unique best hit: 10439073 Mapping efficiency: 40.1% Sequence pairs with no alignments under any condition: 13324084 Sequence pairs did not map uniquely: 2246240 Sequence pairs which were discarded because genomic sequence could not be extracted: 2 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 5162472 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 5276599 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 313246619 Total methylated C's in CpG context: 12166335 Total methylated C's in CHG context: 1553054 Total methylated C's in CHH context: 3545799 Total methylated C's in Unknown context: 59307 Total unmethylated C's in CpG context: 33102077 Total unmethylated C's in CHG context: 67300222 Total unmethylated C's in CHH context: 195579132 Total unmethylated C's in Unknown context: 676779 C methylated in CpG context: 26.9% C methylated in CHG context: 2.3% C methylated in CHH context: 1.8% C methylated in unknown context (CN or CHN): 8.1% Bismark completed in 0d 1h 41m 2s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-154_S5_L002_R1_001_val_1.fq.gz to EPI-154_S5_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-154_S5_L002_R1_001_val_1.fq.gz (25226760 sequences in total) Writing a G -> A converted version of the input file EPI-154_S5_L002_R2_001_val_2.fq.gz to EPI-154_S5_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-154_S5_L002_R2_001_val_2.fq.gz (25226760 sequences in total) Input files are EPI-154_S5_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-154_S5_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-154_S5_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-154_S5_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:2191:2242_1:N:0:TGACCA/1 77 * 0 0 * * 0 0 ATGTANATTAAGAGTTAAATTATGTAGATGTAGATTAAAAGTTAAATTATGTATAGGTAGATTAAGAGTTAAATTAAGTAGATGTAGATTAAGAGTTTAAT BBBBB#>> Writing bisulfite mapping results to EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far Processed 23000000 sequence pairs so far Processed 24000000 sequence pairs so far Processed 25000000 sequence pairs so far 2522676025226760 reads; of these: reads; of these: 2522676025226760 ( (100.00100.00%%) were paired; of these:) were paired; of these: 1756118917487663 ( (69.6169.32%%) aligned concordantly 0 times) aligned concordantly 0 times 34445573441962 ( (13.6513.64%%) aligned concordantly exactly 1 time) aligned concordantly exactly 1 time 42210144297135 ( (16.7317.03%%) aligned concordantly >1 times) aligned concordantly >1 times 30.3930.68%% overall alignment rate overall alignment rate Processed 25226760 sequences in total Successfully deleted the temporary files EPI-154_S5_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-154_S5_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 25226760 Number of paired-end alignments with a unique best hit: 9405544 Mapping efficiency: 37.3% Sequence pairs with no alignments under any condition: 13904236 Sequence pairs did not map uniquely: 1916980 Sequence pairs which were discarded because genomic sequence could not be extracted: 0 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 4637602 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 4767942 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 285373397 Total methylated C's in CpG context: 11306006 Total methylated C's in CHG context: 1298778 Total methylated C's in CHH context: 3449101 Total methylated C's in Unknown context: 62682 Total unmethylated C's in CpG context: 28242299 Total unmethylated C's in CHG context: 60128621 Total unmethylated C's in CHH context: 180948592 Total unmethylated C's in Unknown context: 625876 C methylated in CpG context: 28.6% C methylated in CHG context: 2.1% C methylated in CHH context: 1.9% C methylated in unknown context (CN or CHN): 9.1% Bismark completed in 0d 1h 32m 16s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-159_S6_L002_R1_001_val_1.fq.gz to EPI-159_S6_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-159_S6_L002_R1_001_val_1.fq.gz (15453987 sequences in total) Writing a G -> A converted version of the input file EPI-159_S6_L002_R2_001_val_2.fq.gz to EPI-159_S6_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-159_S6_L002_R2_001_val_2.fq.gz (15453987 sequences in total) Input files are EPI-159_S6_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-159_S6_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-159_S6_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-159_S6_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:1469:2230_1:N:0:ACAGTG/1 77 * 0 0 * * 0 0 GGTTANTGAAGTGTATATAGTTATATTGTAAATTAATGGTTGAGTGAGTTTTNTTTTTTATTAGGTAATATTTTAAGGGAATGGTTGTTTGTAAATATTGG BBBBB#BBFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF/FFFFFFF#<>> Writing bisulfite mapping results to EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far 15453987 reads; of these: 15453987 (100.00%) were paired; of these: 10169665 (65.81%) aligned concordantly 0 times 2347488 (15.19%) aligned concordantly exactly 1 time 2936834 (19.00%) aligned concordantly >1 times 34.19% overall alignment rate 15453987 reads; of these: 15453987 (100.00%) were paired; of these: 10221296 (66.14%) aligned concordantly 0 times 2334749 (15.11%) aligned concordantly exactly 1 time 2897942 (18.75%) aligned concordantly >1 times 33.86% overall alignment rate Processed 15453987 sequences in total Successfully deleted the temporary files EPI-159_S6_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-159_S6_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 15453987 Number of paired-end alignments with a unique best hit: 6380873 Mapping efficiency: 41.3% Sequence pairs with no alignments under any condition: 7739907 Sequence pairs did not map uniquely: 1333207 Sequence pairs which were discarded because genomic sequence could not be extracted: 0 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 3152381 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 3228492 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 195385696 Total methylated C's in CpG context: 7804592 Total methylated C's in CHG context: 1021997 Total methylated C's in CHH context: 3443697 Total methylated C's in Unknown context: 54626 Total unmethylated C's in CpG context: 19140558 Total unmethylated C's in CHG context: 40545239 Total unmethylated C's in CHH context: 123429613 Total unmethylated C's in Unknown context: 431888 C methylated in CpG context: 29.0% C methylated in CHG context: 2.5% C methylated in CHH context: 2.7% C methylated in unknown context (CN or CHN): 11.2% Bismark completed in 0d 1h 1m 41s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-160_S7_L002_R1_001_val_1.fq.gz to EPI-160_S7_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-160_S7_L002_R1_001_val_1.fq.gz (31922572 sequences in total) Writing a G -> A converted version of the input file EPI-160_S7_L002_R2_001_val_2.fq.gz to EPI-160_S7_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-160_S7_L002_R2_001_val_2.fq.gz (31922572 sequences in total) Input files are EPI-160_S7_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-160_S7_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-160_S7_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-160_S7_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:1522:2231_1:N:0:GCCAAT/1 77 * 0 0 * * 0 0 GTATGNGTAATTAGGTGATTTTTTTTGATTTGATGTAGTTAGTGGATTGGGAATTGGTAA BBBBB#BBBFFBFFFFFFFFFFFFFFB/>> Writing bisulfite mapping results to EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:1114:12011:63361_1:N:0:GCCAAT PGA_scaffold16__33_contigs__length_31980953 2 Processed 23000000 sequence pairs so far Processed 24000000 sequence pairs so far Processed 25000000 sequence pairs so far Processed 26000000 sequence pairs so far Processed 27000000 sequence pairs so far Processed 28000000 sequence pairs so far Processed 29000000 sequence pairs so far Processed 30000000 sequence pairs so far Processed 31000000 sequence pairs so far 31922572 reads; of these: 31922572 (100.00%) were paired; of these: 21498211 (67.34%) aligned concordantly 0 times 4482845 (14.04%) aligned concordantly exactly 1 time 5941516 (18.61%) aligned concordantly >1 times 32.66% overall alignment rate 31922572 reads; of these: 31922572 (100.00%) were paired; of these: 21367255 (66.93%) aligned concordantly 0 times 4496752 (14.09%) aligned concordantly exactly 1 time 6058565 (18.98%) aligned concordantly >1 times 33.07% overall alignment rate Processed 31922572 sequences in total Successfully deleted the temporary files EPI-160_S7_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-160_S7_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 31922572 Number of paired-end alignments with a unique best hit: 12255649 Mapping efficiency: 38.4% Sequence pairs with no alignments under any condition: 16675995 Sequence pairs did not map uniquely: 2990928 Sequence pairs which were discarded because genomic sequence could not be extracted: 1 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 6034016 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 6221632 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 356119849 Total methylated C's in CpG context: 13819770 Total methylated C's in CHG context: 1872039 Total methylated C's in CHH context: 4292886 Total methylated C's in Unknown context: 84309 Total unmethylated C's in CpG context: 37264731 Total unmethylated C's in CHG context: 75867945 Total unmethylated C's in CHH context: 223002478 Total unmethylated C's in Unknown context: 787408 C methylated in CpG context: 27.1% C methylated in CHG context: 2.4% C methylated in CHH context: 1.9% C methylated in unknown context (CN or CHN): 9.7% Bismark completed in 0d 1h 57m 46s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-161_S8_L002_R1_001_val_1.fq.gz to EPI-161_S8_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-161_S8_L002_R1_001_val_1.fq.gz (24510805 sequences in total) Writing a G -> A converted version of the input file EPI-161_S8_L002_R2_001_val_2.fq.gz to EPI-161_S8_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-161_S8_L002_R2_001_val_2.fq.gz (24510805 sequences in total) Input files are EPI-161_S8_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-161_S8_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-161_S8_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-161_S8_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:2010:2229_1:N:0:CAGATC/1 77 * 0 0 * * 0 0 TGATANGTAGTATATGGTGTATGATATAGTA BBBBB#>> Writing bisulfite mapping results to EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:2114:19616:68633_1:N:0:CAGATC PGA_scaffold16__33_contigs__length_31980953 1 Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:1112:4795:6211_1:N:0:CAGATC PGA_scaffold16__33_contigs__length_31980953 2 Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far Processed 23000000 sequence pairs so far Processed 24000000 sequence pairs so far 24510805 reads; of these: 24510805 (100.00%) were paired; of these: 17297281 (70.57%) aligned concordantly 0 times 3166248 (12.92%) aligned concordantly exactly 1 time 4047276 (16.51%) aligned concordantly >1 times 29.43% overall alignment rate 24510805 reads; of these: 24510805 (100.00%) were paired; of these: 17217869 (70.25%) aligned concordantly 0 times 3175317 (12.95%) aligned concordantly exactly 1 time 4117619 (16.80%) aligned concordantly >1 times 29.75% overall alignment rate Processed 24510805 sequences in total Successfully deleted the temporary files EPI-161_S8_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-161_S8_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 24510805 Number of paired-end alignments with a unique best hit: 8593707 Mapping efficiency: 35.1% Sequence pairs with no alignments under any condition: 13929047 Sequence pairs did not map uniquely: 1988051 Sequence pairs which were discarded because genomic sequence could not be extracted: 2 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 4237007 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 4356698 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 251244640 Total methylated C's in CpG context: 9311185 Total methylated C's in CHG context: 1241250 Total methylated C's in CHH context: 3334595 Total methylated C's in Unknown context: 63649 Total unmethylated C's in CpG context: 25878911 Total unmethylated C's in CHG context: 52710364 Total unmethylated C's in CHH context: 158768335 Total unmethylated C's in Unknown context: 546831 C methylated in CpG context: 26.5% C methylated in CHG context: 2.3% C methylated in CHH context: 2.1% C methylated in unknown context (CN or CHN): 10.4% Bismark completed in 0d 1h 24m 55s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-162_S9_L002_R1_001_val_1.fq.gz to EPI-162_S9_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-162_S9_L002_R1_001_val_1.fq.gz (22769546 sequences in total) Writing a G -> A converted version of the input file EPI-162_S9_L002_R2_001_val_2.fq.gz to EPI-162_S9_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-162_S9_L002_R2_001_val_2.fq.gz (22769546 sequences in total) Input files are EPI-162_S9_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-162_S9_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-162_S9_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-162_S9_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:1238:2233_1:N:0:ACTTGA/1 77 * 0 0 * * 0 0 TGTTANTTATTTTAAAATTTGTTGATAAATTGTTTATTTTTAGGATAGTTTTTTTAGATATTGTTTTTTTTTGGTTTGAAAGGGTTATGAATAGTTTTTTT BBBBB#>> Writing bisulfite mapping results to EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far 22769546 reads; of these: 22769546 (100.00%) were paired; of these: 15681883 (68.87%) aligned concordantly 0 times 3024375 (13.28%) aligned concordantly exactly 1 time 4063288 (17.85%) aligned concordantly >1 times 31.13% overall alignment rate 22769546 reads; of these: 22769546 (100.00%) were paired; of these: 15616786 (68.59%) aligned concordantly 0 times 3009100 (13.22%) aligned concordantly exactly 1 time 4143660 (18.20%) aligned concordantly >1 times 31.41% overall alignment rate Processed 22769546 sequences in total Successfully deleted the temporary files EPI-162_S9_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-162_S9_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 22769546 Number of paired-end alignments with a unique best hit: 8267039 Mapping efficiency: 36.3% Sequence pairs with no alignments under any condition: 12467371 Sequence pairs did not map uniquely: 2035136 Sequence pairs which were discarded because genomic sequence could not be extracted: 0 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 4073674 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 4193365 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 238005650 Total methylated C's in CpG context: 8810888 Total methylated C's in CHG context: 1190010 Total methylated C's in CHH context: 3285737 Total methylated C's in Unknown context: 56519 Total unmethylated C's in CpG context: 24654378 Total unmethylated C's in CHG context: 50555362 Total unmethylated C's in CHH context: 149509275 Total unmethylated C's in Unknown context: 528637 C methylated in CpG context: 26.3% C methylated in CHG context: 2.3% C methylated in CHH context: 2.2% C methylated in unknown context (CN or CHN): 9.7% Bismark completed in 0d 1h 23m 10s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-167_S10_L002_R1_001_val_1.fq.gz to EPI-167_S10_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-167_S10_L002_R1_001_val_1.fq.gz (24481250 sequences in total) Writing a G -> A converted version of the input file EPI-167_S10_L002_R2_001_val_2.fq.gz to EPI-167_S10_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-167_S10_L002_R2_001_val_2.fq.gz (24481250 sequences in total) Input files are EPI-167_S10_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-167_S10_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-167_S10_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-167_S10_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:1666:2232_1:N:0:GATCAG/1 77 * 0 0 * * 0 0 TTGAANTTGTTGATTTATATGGGGATATGAATAATAAGTTTAATATTTAAGATGGAGAAGAAATAAGTGTAGAATTTTGGATAAAGGATAATTAAGTTAAT BBBBB#BBFFFFFF>> Writing bisulfite mapping results to EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far Processed 23000000 sequence pairs so far Processed 24000000 sequence pairs so far 24481250 reads; of these: 24481250 (100.00%) were paired; of these: 16050656 (65.56%) aligned concordantly 0 times 3801758 (15.53%) aligned concordantly exactly 1 time 4628836 (18.91%) aligned concordantly >1 times 34.44% overall alignment rate 24481250 reads; of these: 24481250 (100.00%) were paired; of these: 16118278 (65.84%) aligned concordantly 0 times 3811968 (15.57%) aligned concordantly exactly 1 time 4551004 (18.59%) aligned concordantly >1 times 34.16% overall alignment rate Processed 24481250 sequences in total Successfully deleted the temporary files EPI-167_S10_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-167_S10_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 24481250 Number of paired-end alignments with a unique best hit: 10438000 Mapping efficiency: 42.6% Sequence pairs with no alignments under any condition: 12046563 Sequence pairs did not map uniquely: 1996687 Sequence pairs which were discarded because genomic sequence could not be extracted: 0 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 5166645 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 5271355 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 327276929 Total methylated C's in CpG context: 12357096 Total methylated C's in CHG context: 2047643 Total methylated C's in CHH context: 6308688 Total methylated C's in Unknown context: 65277 Total unmethylated C's in CpG context: 31908920 Total unmethylated C's in CHG context: 67977563 Total unmethylated C's in CHH context: 206677019 Total unmethylated C's in Unknown context: 705078 C methylated in CpG context: 27.9% C methylated in CHG context: 2.9% C methylated in CHH context: 3.0% C methylated in unknown context (CN or CHN): 8.5% Bismark completed in 0d 1h 40m 24s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-168_S11_L002_R1_001_val_1.fq.gz to EPI-168_S11_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-168_S11_L002_R1_001_val_1.fq.gz (22688467 sequences in total) Writing a G -> A converted version of the input file EPI-168_S11_L002_R2_001_val_2.fq.gz to EPI-168_S11_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-168_S11_L002_R2_001_val_2.fq.gz (22688467 sequences in total) Input files are EPI-168_S11_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-168_S11_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-168_S11_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-168_S11_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:1424:2239_1:N:0:TAGCTT/1 77 * 0 0 * * 0 0 GTGATNTATAATGTATTATTGAAAGTTAATAGTTTATTTAAGTTTTTGGTATTTTTTGATTTAGGTTTAATTATTTTGTTATGTGGGTTTGTGATTTG BBBBB#B>> Writing bisulfite mapping results to EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far 22688467 reads; of these: 22688467 (100.00%) were paired; of these: 15440681 (68.06%) aligned concordantly 0 times 3190562 (14.06%) aligned concordantly exactly 1 time 4057224 (17.88%) aligned concordantly >1 times 31.94% overall alignment rate 22688467 reads; of these: 22688467 (100.00%) were paired; of these: 15321747 (67.53%) aligned concordantly 0 times 3226165 (14.22%) aligned concordantly exactly 1 time 4140555 (18.25%) aligned concordantly >1 times 32.47% overall alignment rate Processed 22688467 sequences in total Successfully deleted the temporary files EPI-168_S11_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-168_S11_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 22688467 Number of paired-end alignments with a unique best hit: 8967128 Mapping efficiency: 39.5% Sequence pairs with no alignments under any condition: 11891374 Sequence pairs did not map uniquely: 1829965 Sequence pairs which were discarded because genomic sequence could not be extracted: 0 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 4393790 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 4573338 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 278794431 Total methylated C's in CpG context: 10658385 Total methylated C's in CHG context: 1268483 Total methylated C's in CHH context: 3535297 Total methylated C's in Unknown context: 54743 Total unmethylated C's in CpG context: 29012483 Total unmethylated C's in CHG context: 59889529 Total unmethylated C's in CHH context: 174430254 Total unmethylated C's in Unknown context: 615881 C methylated in CpG context: 26.9% C methylated in CHG context: 2.1% C methylated in CHH context: 2.0% C methylated in unknown context (CN or CHN): 8.2% Bismark completed in 0d 1h 29m 25s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-169_S12_L002_R1_001_val_1.fq.gz to EPI-169_S12_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-169_S12_L002_R1_001_val_1.fq.gz (21392200 sequences in total) Writing a G -> A converted version of the input file EPI-169_S12_L002_R2_001_val_2.fq.gz to EPI-169_S12_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-169_S12_L002_R2_001_val_2.fq.gz (21392200 sequences in total) Input files are EPI-169_S12_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-169_S12_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-169_S12_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-169_S12_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:1452:2231_1:N:0:GGCTAC/1 77 * 0 0 * * 0 0 TGGTANGAGGTTGTGTGATTAGTATAAAGGTGTTTAGAGTTATTAAGTGGATTGGATTGGAGTTTGGATTGGTTTTGTATTAATAAAAGTGTTTTTTTTGT BBBBB#<>> Writing bisulfite mapping results to EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R2_001_val_2.fq.gz Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:2205:7072:26521_1:N:0:GGCTAC PGA_scaffold9__45_contigs__length_38581958 1 Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:1306:7118:3504_1:N:0:GGCTAC PGA_scaffold16__33_contigs__length_31980953 2 Processed 11000000 sequence pairs so far Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:1208:9887:13727_1:N:0:GGCTAC PGA_scaffold16__33_contigs__length_31980953 2 Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:1114:15881:53162_1:N:0:GGCTAC PGA_scaffold10__49_contigs__length_53961475 1 Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far 21392200 reads; of these: 21392200 (100.00%) were paired; of these: 14178782 (66.28%) aligned concordantly 0 times 3054776 (14.28%) aligned concordantly exactly 1 time 4158642 (19.44%) aligned concordantly >1 times 33.72% overall alignment rate 21392200 reads; of these: 21392200 (100.00%) were paired; of these: 14275451 (66.73%) aligned concordantly 0 times 3039394 (14.21%) aligned concordantly exactly 1 time 4077355 (19.06%) aligned concordantly >1 times 33.27% overall alignment rate Processed 21392200 sequences in total Successfully deleted the temporary files EPI-169_S12_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-169_S12_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 21392200 Number of paired-end alignments with a unique best hit: 8408059 Mapping efficiency: 39.3% Sequence pairs with no alignments under any condition: 10932504 Sequence pairs did not map uniquely: 2051637 Sequence pairs which were discarded because genomic sequence could not be extracted: 4 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 4126394 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 4281661 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 245148711 Total methylated C's in CpG context: 9715161 Total methylated C's in CHG context: 1293014 Total methylated C's in CHH context: 3765782 Total methylated C's in Unknown context: 64209 Total unmethylated C's in CpG context: 25600954 Total unmethylated C's in CHG context: 52728521 Total unmethylated C's in CHH context: 152045279 Total unmethylated C's in Unknown context: 533680 C methylated in CpG context: 27.5% C methylated in CHG context: 2.4% C methylated in CHH context: 2.4% C methylated in unknown context (CN or CHN): 10.7% Bismark completed in 0d 1h 21m 2s ==================== Bismark run complete ==================== Use of uninitialized value $path_to_bowtie in concatenation (.) or string at /gscratch/srlab/programs/Bismark-0.21.0/bismark line 6893. Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.1.0]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/gscratch/srlab/programs/samtools-1.4/bin/samtools' Reference genome folder provided is /gscratch/srlab/sr320/data/geoduck/v074/ (absolute path is '/gscratch/srlab/sr320/data/geoduck/v074/)' FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 4 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/gscratch/scrubbed/sr320/0807'): /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R1_001_val_1.fq.gz /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R2_001_val_2.fq.gz Library is assumed to be strand-specific (directional), alignments to strands complementary to the original top or bottom strands will be ignored (i.e. not performed!) Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /gscratch/scrubbed/sr320/0807 Now reading in and storing sequence information of the genome specified in: /gscratch/srlab/sr320/data/geoduck/v074/ chr PGA_scaffold1__77_contigs__length_89643857 (89643857 bp) chr PGA_scaffold2__36_contigs__length_69596280 (69596280 bp) chr PGA_scaffold3__111_contigs__length_57743597 (57743597 bp) chr PGA_scaffold4__129_contigs__length_65288255 (65288255 bp) chr PGA_scaffold5__109_contigs__length_67248332 (67248332 bp) chr PGA_scaffold6__104_contigs__length_61759565 (61759565 bp) chr PGA_scaffold7__69_contigs__length_43120122 (43120122 bp) chr PGA_scaffold8__63_contigs__length_61151155 (61151155 bp) chr PGA_scaffold9__45_contigs__length_38581958 (38581958 bp) chr PGA_scaffold10__49_contigs__length_53961475 (53961475 bp) chr PGA_scaffold11__79_contigs__length_51449921 (51449921 bp) chr PGA_scaffold12__71_contigs__length_50438331 (50438331 bp) chr PGA_scaffold13__52_contigs__length_44396874 (44396874 bp) chr PGA_scaffold14__91_contigs__length_45393038 (45393038 bp) chr PGA_scaffold15__101_contigs__length_47938513 (47938513 bp) chr PGA_scaffold16__33_contigs__length_31980953 (31980953 bp) chr PGA_scaffold17__51_contigs__length_34923512 (34923512 bp) chr PGA_scaffold18__69_contigs__length_27737463 (27737463 bp) Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R2_001_val_2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file EPI-170_S13_L002_R1_001_val_1.fq.gz to EPI-170_S13_L002_R1_001_val_1.fq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file EPI-170_S13_L002_R1_001_val_1.fq.gz (27996279 sequences in total) Writing a G -> A converted version of the input file EPI-170_S13_L002_R2_001_val_2.fq.gz to EPI-170_S13_L002_R2_001_val_2.fq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file EPI-170_S13_L002_R2_001_val_2.fq.gz (27996279 sequences in total) Input files are EPI-170_S13_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-170_S13_L002_R2_001_val_2.fq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /gscratch/srlab/sr320/data/geoduck/v074/ with the specified options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from EPI-170_S13_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-170_S13_L002_R2_001_val_2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.6 -p 4 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: D00743:144:CAAWNANXX:2:2207:2244:2228_1:N:0:CTTGTA/1 77 * 0 0 * * 0 0 TTGTTNAGTTTGTTTAAGTTATTTTGATTTGTTAAAAAATATGGTAG BBBBB#<>> Writing bisulfite mapping results to EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.bam <<< Reading in the sequence files /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R1_001_val_1.fq.gz and /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R2_001_val_2.fq.gz Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:2107:18595:54334_1:N:0:CTTGTA PGA_scaffold16__33_contigs__length_31980953 2 Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far Processed 6000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:2111:5981:74801_1:N:0:CTTGTA PGA_scaffold16__33_contigs__length_31980953 3 Processed 7000000 sequence pairs so far Processed 8000000 sequence pairs so far Processed 9000000 sequence pairs so far Processed 10000000 sequence pairs so far Processed 11000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:1316:10202:11032_1:N:0:CTTGTA PGA_scaffold16__33_contigs__length_31980953 2 Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:1314:12592:63283_1:N:0:CTTGTA PGA_scaffold15__101_contigs__length_47938513 3 Processed 12000000 sequence pairs so far Processed 13000000 sequence pairs so far Processed 14000000 sequence pairs so far Processed 15000000 sequence pairs so far Processed 16000000 sequence pairs so far Processed 17000000 sequence pairs so far Processed 18000000 sequence pairs so far Processed 19000000 sequence pairs so far Processed 20000000 sequence pairs so far Processed 21000000 sequence pairs so far Processed 22000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:2213:8607:54584_1:N:0:CTTGTA PGA_scaffold9__45_contigs__length_38581958 2 Processed 23000000 sequence pairs so far Processed 24000000 sequence pairs so far Processed 25000000 sequence pairs so far Processed 26000000 sequence pairs so far Processed 27000000 sequence pairs so far Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:2312:8356:27499_1:N:0:CTTGTA PGA_scaffold15__101_contigs__length_47938513 3 Chromosomal sequence could not be extracted for D00743:144:CAAWNANXX:2:2312:6036:58634_1:N:0:CTTGTA PGA_scaffold15__101_contigs__length_47938513 3 27996279 reads; of these: 27996279 (100.00%) were paired; of these: 19276145 (68.85%) aligned concordantly 0 times 3764638 (13.45%) aligned concordantly exactly 1 time 4955496 (17.70%) aligned concordantly >1 times 31.15% overall alignment rate 27996279 reads; of these: 27996279 (100.00%) were paired; of these: 19163557 (68.45%) aligned concordantly 0 times 3776883 (13.49%) aligned concordantly exactly 1 time 5055839 (18.06%) aligned concordantly >1 times 31.55% overall alignment rate Processed 27996279 sequences in total Successfully deleted the temporary files EPI-170_S13_L002_R1_001_val_1.fq.gz_C_to_T.fastq and EPI-170_S13_L002_R2_001_val_2.fq.gz_G_to_A.fastq Final Alignment report ====================== Sequence pairs analysed in total: 27996279 Number of paired-end alignments with a unique best hit: 10233767 Mapping efficiency: 36.6% Sequence pairs with no alignments under any condition: 15243065 Sequence pairs did not map uniquely: 2519447 Sequence pairs which were discarded because genomic sequence could not be extracted: 7 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 5017772 ((converted) top strand) GA/CT/CT: 0 (complementary to (converted) top strand) GA/CT/GA: 0 (complementary to (converted) bottom strand) CT/GA/GA: 5215988 ((converted) bottom strand) Number of alignments to (merely theoretical) complementary strands being rejected in total: 0 Final Cytosine Methylation Report ================================= Total number of C's analysed: 295230497 Total methylated C's in CpG context: 10743620 Total methylated C's in CHG context: 1476561 Total methylated C's in CHH context: 3272606 Total methylated C's in Unknown context: 54419 Total unmethylated C's in CpG context: 31907626 Total unmethylated C's in CHG context: 62815831 Total unmethylated C's in CHH context: 185014253 Total unmethylated C's in Unknown context: 638215 C methylated in CpG context: 25.2% C methylated in CHG context: 2.3% C methylated in CHH context: 1.7% C methylated in unknown context (CN or CHN): 7.9% Bismark completed in 0d 1h 39m 24s ==================== Bismark run complete ==================== Processing paired-end Bismark output file(s) (SAM format): EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.bam: 9915441 Total number duplicated alignments removed: 2705164 (27.28%) Duplicated alignments were found at: 1698251 different position(s) Total count of deduplicated leftover sequences: 7210277 (72.72% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.bam: 11727816 Total number duplicated alignments removed: 3318046 (28.29%) Duplicated alignments were found at: 2042936 different position(s) Total count of deduplicated leftover sequences: 8409770 (71.71% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.bam: 10439071 Total number duplicated alignments removed: 3739516 (35.82%) Duplicated alignments were found at: 2094280 different position(s) Total count of deduplicated leftover sequences: 6699555 (64.18% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.bam: 9405544 Total number duplicated alignments removed: 2636021 (28.03%) Duplicated alignments were found at: 1665648 different position(s) Total count of deduplicated leftover sequences: 6769523 (71.97% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.bam: 6380873 Total number duplicated alignments removed: 1736608 (27.22%) Duplicated alignments were found at: 1097543 different position(s) Total count of deduplicated leftover sequences: 4644265 (72.78% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.bam: 12255648 Total number duplicated alignments removed: 3263582 (26.63%) Duplicated alignments were found at: 1915571 different position(s) Total count of deduplicated leftover sequences: 8992066 (73.37% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.bam: 8593705 Total number duplicated alignments removed: 2262879 (26.33%) Duplicated alignments were found at: 1443182 different position(s) Total count of deduplicated leftover sequences: 6330826 (73.67% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.bam: 8267039 Total number duplicated alignments removed: 1883875 (22.79%) Duplicated alignments were found at: 1160963 different position(s) Total count of deduplicated leftover sequences: 6383164 (77.21% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.bam: 10438000 Total number duplicated alignments removed: 2802912 (26.85%) Duplicated alignments were found at: 1823622 different position(s) Total count of deduplicated leftover sequences: 7635088 (73.15% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.bam: 8967128 Total number duplicated alignments removed: 2474742 (27.60%) Duplicated alignments were found at: 1499379 different position(s) Total count of deduplicated leftover sequences: 6492386 (72.40% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.bam: 8408055 Total number duplicated alignments removed: 1984624 (23.60%) Duplicated alignments were found at: 1201335 different position(s) Total count of deduplicated leftover sequences: 6423431 (76.40% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R2_001_val_2.fq.gz" Processing paired-end Bismark output file(s) (SAM format): EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.bam: 10233760 Total number duplicated alignments removed: 2436682 (23.81%) Duplicated alignments were found at: 1417870 different position(s) Total count of deduplicated leftover sequences: 7797078 (76.19% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R2_001_val_2.fq.gz" *** Bismark methylation extractor version v0.21.0 *** Trying to determine the type of mapping from the SAM header line of file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Treating file(s) as paired-end data (as extracted from @PG line) Setting option '--no_overlap' since this is (normally) the right thing to do for paired-end data Summarising Bismark methylation extractor parameters: =============================================================== Bismark paired-end SAM format specified (default) Number of cores to be used: 14 Output will be written to the current directory ('/gscratch/scrubbed/sr320/0807') Summarising bedGraph parameters: =============================================================== Generating additional output in bedGraph and coverage format bedGraph format: coverage format: Using a cutoff of 1 read(s) to report cytosine positions Reporting and sorting cytosine methylation information in CpG context only (default) The bedGraph UNIX sort command will use the following memory setting: '75%'. Temporary directory used for sorting is the output directory Checking file >>EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-151_S2_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 1500000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2000000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 2500000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3000000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 3500000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5000000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 7210277 lines in total Total number of methylation call strings processed: 14420554 Final Cytosine Methylation Report ================================= Total number of C's analysed: 154371720 Total methylated C's in CpG context: 6619617 Total methylated C's in CHG context: 739999 Total methylated C's in CHH context: 1943298 Total C to T conversions in CpG context: 14863708 Total C to T conversions in CHG context: 32156644 Total C to T conversions in CHH context: 98048454 C methylated in CpG context: 30.8% C methylated in CHG context: 2.2% C methylated in CHH context: 1.9% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-152_S3_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7000000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 7500000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 8000000 Processed lines: 8000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 8000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 8409770 lines in total Total number of methylation call strings processed: 16819540 Final Cytosine Methylation Report ================================= Total number of C's analysed: 181742375 Total methylated C's in CpG context: 7874020 Total methylated C's in CHG context: 929405 Total methylated C's in CHH context: 2251146 Total C to T conversions in CpG context: 16740253 Total C to T conversions in CHG context: 36920601 Total C to T conversions in CHH context: 117026950 C methylated in CpG context: 32.0% C methylated in CHG context: 2.5% C methylated in CHH context: 1.9% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-153_S4_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 6000000 Processed lines: 5500000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6500000 Processed lines: 6000000 Finished processing child process. Exiting.. Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Processed lines: 6500000 Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 6699555 lines in total Total number of methylation call strings processed: 13399110 Final Cytosine Methylation Report ================================= Total number of C's analysed: 136016129 Total methylated C's in CpG context: 5750945 Total methylated C's in CHG context: 699624 Total methylated C's in CHH context: 1733351 Total C to T conversions in CpG context: 13021245 Total C to T conversions in CHG context: 28351144 Total C to T conversions in CHH context: 86459820 C methylated in CpG context: 30.6% C methylated in CHG context: 2.4% C methylated in CHH context: 2.0% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-154_S5_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Processed lines: 6500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 6500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 6769523 lines in total Total number of methylation call strings processed: 13539046 Final Cytosine Methylation Report ================================= Total number of C's analysed: 142831749 Total methylated C's in CpG context: 5958725 Total methylated C's in CHG context: 672814 Total methylated C's in CHH context: 1876297 Total C to T conversions in CpG context: 13541358 Total C to T conversions in CHG context: 29209282 Total C to T conversions in CHH context: 91573273 C methylated in CpG context: 30.6% C methylated in CHG context: 2.3% C methylated in CHH context: 2.0% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-159_S6_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Processed lines: 4500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 4500000 Finished processing child process. Exiting.. Now waiting for all child processes to complete Merging individual splitting reports into overall report: 'EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 4644265 lines in total Total number of methylation call strings processed: 9288530 Final Cytosine Methylation Report ================================= Total number of C's analysed: 99137282 Total methylated C's in CpG context: 4217985 Total methylated C's in CHG context: 535634 Total methylated C's in CHH context: 1932988 Total C to T conversions in CpG context: 9196803 Total C to T conversions in CHG context: 20037165 Total C to T conversions in CHH context: 63216707 C methylated in CpG context: 31.4% C methylated in CHG context: 2.6% C methylated in CHH context: 3.0% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-160_S7_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 8500000 Processed lines: 8500000 Processed lines: 8500000 Processed lines: 8500000 Processed lines: 8500000 Processed lines: 8500000 Processed lines: 8500000 Finished processing child process. Exiting.. Processed lines: 8500000 Processed lines: 8500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 8992066 lines in total Total number of methylation call strings processed: 17984132 Final Cytosine Methylation Report ================================= Total number of C's analysed: 176538868 Total methylated C's in CpG context: 7344361 Total methylated C's in CHG context: 912079 Total methylated C's in CHH context: 2297548 Total C to T conversions in CpG context: 16933790 Total C to T conversions in CHG context: 36262599 Total C to T conversions in CHH context: 112788491 C methylated in CpG context: 30.3% C methylated in CHG context: 2.5% C methylated in CHH context: 2.0% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-161_S8_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Finished processing child process. Exiting.. Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 6330826 lines in total Total number of methylation call strings processed: 12661652 Final Cytosine Methylation Report ================================= Total number of C's analysed: 124508835 Total methylated C's in CpG context: 4903316 Total methylated C's in CHG context: 619102 Total methylated C's in CHH context: 1795108 Total C to T conversions in CpG context: 12097133 Total C to T conversions in CHG context: 25334277 Total C to T conversions in CHH context: 79759899 C methylated in CpG context: 28.8% C methylated in CHG context: 2.4% C methylated in CHH context: 2.2% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-162_S9_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Merging individual splitting reports into overall report: 'EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 6383164 lines in total Total number of methylation call strings processed: 12766328 Final Cytosine Methylation Report ================================= Total number of C's analysed: 122908719 Total methylated C's in CpG context: 4824979 Total methylated C's in CHG context: 612515 Total methylated C's in CHH context: 1827593 Total C to T conversions in CpG context: 12090566 Total C to T conversions in CHG context: 25238601 Total C to T conversions in CHH context: 78314465 C methylated in CpG context: 28.5% C methylated in CHG context: 2.4% C methylated in CHH context: 2.3% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-167_S10_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7500000 Finished processing child process. Exiting.. Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Finished processing child process. Exiting.. Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Finished processing child process. Exiting.. Processed lines: 7500000 Finished processing child process. Exiting.. Processed lines: 7500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 7635088 lines in total Total number of methylation call strings processed: 15270176 Final Cytosine Methylation Report ================================= Total number of C's analysed: 169475057 Total methylated C's in CpG context: 6721143 Total methylated C's in CHG context: 1081552 Total methylated C's in CHH context: 3482873 Total C to T conversions in CpG context: 15986854 Total C to T conversions in CHG context: 34252524 Total C to T conversions in CHH context: 107950111 C methylated in CpG context: 29.6% C methylated in CHG context: 3.1% C methylated in CHH context: 3.1% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-168_S11_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 6492386 lines in total Total number of methylation call strings processed: 12984772 Final Cytosine Methylation Report ================================= Total number of C's analysed: 139836026 Total methylated C's in CpG context: 5749134 Total methylated C's in CHG context: 652679 Total methylated C's in CHH context: 1958750 Total C to T conversions in CpG context: 13380784 Total C to T conversions in CHG context: 28960958 Total C to T conversions in CHH context: 89133721 C methylated in CpG context: 30.1% C methylated in CHG context: 2.2% C methylated in CHH context: 2.2% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-169_S12_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 5500000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Processed lines: 6000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 6423431 lines in total Total number of methylation call strings processed: 12846862 Final Cytosine Methylation Report ================================= Total number of C's analysed: 127237948 Total methylated C's in CpG context: 5351229 Total methylated C's in CHG context: 663359 Total methylated C's in CHH context: 2095978 Total C to T conversions in CpG context: 12254820 Total C to T conversions in CHG context: 26439097 Total C to T conversions in CHH context: 80433465 C methylated in CpG context: 30.4% C methylated in CHG context: 2.4% C methylated in CHH context: 2.5% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Checking file >>EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam<< for signs of file truncation... Now testing Bismark result file >EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam< for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Now reading in Bismark result file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bam Warning: unable to close filehandle IN properly. skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:PGA_scaffold1__77_contigs__length_89643857 LN:89643857 skipping SAM header line: @SQ SN:PGA_scaffold2__36_contigs__length_69596280 LN:69596280 skipping SAM header line: @SQ SN:PGA_scaffold3__111_contigs__length_57743597 LN:57743597 skipping SAM header line: @SQ SN:PGA_scaffold4__129_contigs__length_65288255 LN:65288255 skipping SAM header line: @SQ SN:PGA_scaffold5__109_contigs__length_67248332 LN:67248332 skipping SAM header line: @SQ SN:PGA_scaffold6__104_contigs__length_61759565 LN:61759565 skipping SAM header line: @SQ SN:PGA_scaffold7__69_contigs__length_43120122 LN:43120122 skipping SAM header line: @SQ SN:PGA_scaffold8__63_contigs__length_61151155 LN:61151155 skipping SAM header line: @SQ SN:PGA_scaffold9__45_contigs__length_38581958 LN:38581958 skipping SAM header line: @SQ SN:PGA_scaffold10__49_contigs__length_53961475 LN:53961475 skipping SAM header line: @SQ SN:PGA_scaffold11__79_contigs__length_51449921 LN:51449921 skipping SAM header line: @SQ SN:PGA_scaffold12__71_contigs__length_50438331 LN:50438331 skipping SAM header line: @SQ SN:PGA_scaffold13__52_contigs__length_44396874 LN:44396874 skipping SAM header line: @SQ SN:PGA_scaffold14__91_contigs__length_45393038 LN:45393038 skipping SAM header line: @SQ SN:PGA_scaffold15__101_contigs__length_47938513 LN:47938513 skipping SAM header line: @SQ SN:PGA_scaffold16__33_contigs__length_31980953 LN:31980953 skipping SAM header line: @SQ SN:PGA_scaffold17__51_contigs__length_34923512 LN:34923512 skipping SAM header line: @SQ SN:PGA_scaffold18__69_contigs__length_27737463 LN:27737463 skipping SAM header line: @PG ID:Bismark VN:v0.21.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ -genome /gscratch/srlab/sr320/data/geoduck/v074 -p 4 -score_min L,0,-0.6 -1 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R1_001_val_1.fq.gz -2 /gscratch/srlab/strigg/data/Pgenr/FASTQS/EPI-170_S13_L002_R2_001_val_2.fq.gz" Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 500000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1000000 Processed lines: 1500000 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Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Now waiting for all child processes to complete Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt' Merging from these individual files: EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13 EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14 Processed 7797078 lines in total Total number of methylation call strings processed: 15594156 Final Cytosine Methylation Report ================================= Total number of C's analysed: 151781122 Total methylated C's in CpG context: 5915753 Total methylated C's in CHG context: 746213 Total methylated C's in CHH context: 1797219 Total C to T conversions in CpG context: 15089108 Total C to T conversions in CHG context: 31172597 Total C to T conversions in CHH context: 97060232 C methylated in CpG context: 28.2% C methylated in CHG context: 2.3% C methylated in CHH context: 1.8% Merging individual M-bias reports into overall M-bias statistics from these 14 individual files: EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.1.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.2.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.3.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.4.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.5.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.6.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.7.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.8.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.9.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.10.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.11.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.12.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.13.mbias EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt.14.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 101 Maximum read length of Read 2: 99 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CpG_CTOT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_CTOB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CpG_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHG_CTOT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_CTOB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHG_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept CHH_CTOT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_CTOB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt was empty -> deleted CHH_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt contains data -> kept Using these input files: CpG_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CpG_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHG_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt CHH_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz output directory: >< remove whitespaces: no CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/sr320/0807/CpG_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt /gscratch/scrubbed/sr320/0807/CpG_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt Writing bedGraph to file: EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished writing methylation calls from CpG_OT_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Finished writing methylation calls from CpG_OB_EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.txt to merged temp file Sorting input file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Found 12 alignment reports in current directory. Now trying to figure out whether there are corresponding optional reports Writing Bismark HTML report to >> EPI-151_S2_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-151_S2_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-151_S2_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-151_S2_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-152_S3_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-152_S3_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-152_S3_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-152_S3_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-153_S4_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-153_S4_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-153_S4_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-153_S4_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-154_S5_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-154_S5_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-154_S5_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-154_S5_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-159_S6_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-159_S6_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-159_S6_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-159_S6_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-160_S7_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-160_S7_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-160_S7_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-160_S7_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-161_S8_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-161_S8_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-161_S8_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-161_S8_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-162_S9_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-162_S9_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-162_S9_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-162_S9_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-167_S10_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-167_S10_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-167_S10_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-167_S10_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-168_S11_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-168_S11_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-168_S11_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-168_S11_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-169_S12_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-169_S12_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-169_S12_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-169_S12_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== Writing Bismark HTML report to >> EPI-170_S13_L002_R1_001_val_1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > EPI-170_S13_L002_R1_001_val_1_bismark_bt2_PE_report.txt < Processing alignment report EPI-170_S13_L002_R1_001_val_1_bismark_bt2_PE_report.txt ... Complete Using the following deduplication report: > EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt < Processing deduplication report EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplication_report.txt ... Complete Using the following splitting report: > EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt < Processing splitting report EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated_splitting_report.txt ... Complete Using the following M-bias report: > EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt < Processing M-bias report EPI-170_S13_L002_R1_001_val_1_bismark_bt2_pe.deduplicated.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== No Bismark/Bowtie2 single-end BAM files detected Found Bismark/Bowtie2 paired-end files No Bismark/HISAT2 single-end BAM files detected No Bismark/HISAT2 paired-end BAM files detected Generating Bismark summary report from 12 Bismark BAM file(s)... >> Reading from Bismark report: EPI-151_S2_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-152_S3_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-153_S4_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-154_S5_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-159_S6_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-160_S7_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-161_S8_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-162_S9_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-167_S10_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-168_S11_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-169_S12_L002_R1_001_val_1_bismark_bt2_PE_report.txt >> Reading from Bismark report: EPI-170_S13_L002_R1_001_val_1_bismark_bt2_PE_report.txt Wrote Bismark project summary to >> bismark_summary_report.html << [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks... [bam_sort_core] merging from 0 files and 28 in-memory blocks...