Bowtie 2 seems to be working fine (tested command 'bowtie2 --version' [2.5.4]) Output format is BAM (default) Alignments will be written out in BAM format. Samtools found here: '/srlab/programs/samtools-1.20/samtools' Reference genome folder provided is /mmfs1/gscratch/scrubbed/sr320/github/Claudia-Halibut/project/data/genome/ (absolute path is '/mmfs1/gscratch/scrubbed/sr320/github/Claudia-Halibut/project/data/genome/)' FastQ format assumed (by default) Attention: early reports suggested that high values of -p to have diminishing returns. Please test different values using a small subset of data for your hardware setting. Each Bowtie 2 instance is going to be run with 8 threads. Please monitor performance closely and tune down if necessary! Input files to be analysed (in current folder '/mmfs1/gscratch/scrubbed/sr320/github/Claudia-Halibut/project/output/02'): /mmfs1/gscratch/scrubbed/sr320/RRBS-run2/raw-data/E2_T1_C1_100_R1.fq.gz /mmfs1/gscratch/scrubbed/sr320/RRBS-run2/raw-data/E2_T1_C1_100_R2.fq.gz Library was specified to be not strand-specific (non-directional), therefore alignments to all four possible bisulfite strands (OT, CTOT, OB and CTOB) will be reported Output will be written into the directory: /mmfs1/gscratch/scrubbed/sr320/github/Claudia-Halibut/project/output/02/ Setting parallelization to single-threaded (default) Summary of all aligner options: -q --score-min L,0,-0.8 -p 8 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Current working directory is: /mmfs1/gscratch/scrubbed/sr320/github/Claudia-Halibut/project/output/02 Now reading in and storing sequence information of the genome specified in: /mmfs1/gscratch/scrubbed/sr320/github/Claudia-Halibut/project/data/genome/ Single-core mode: setting pid to 1 Paired-end alignments will be performed ======================================= The provided filenames for paired-end alignments are /mmfs1/gscratch/scrubbed/sr320/RRBS-run2/raw-data/E2_T1_C1_100_R1.fq.gz and /mmfs1/gscratch/scrubbed/sr320/RRBS-run2/raw-data/E2_T1_C1_100_R2.fq.gz Input files are in FastQ format Writing a C -> T converted version of the input file E2_T1_C1_100_R1.fq.gz to E2_T1_C1_100_R1.fq.gz_C_to_T.fastq Writing a G -> A converted version of the input file E2_T1_C1_100_R1.fq.gz to E2_T1_C1_100_R1.fq.gz_G_to_A.fastq Created C -> T as well as G -> A converted versions of the FastQ file E2_T1_C1_100_R1.fq.gz (86858983 sequences in total) Writing a C -> T converted version of the input file E2_T1_C1_100_R2.fq.gz to E2_T1_C1_100_R2.fq.gz_C_to_T.fastq Writing a G -> A converted version of the input file E2_T1_C1_100_R2.fq.gz to E2_T1_C1_100_R2.fq.gz_G_to_A.fastq Created C -> T as well as G -> A converted versions of the FastQ file E2_T1_C1_100_R2.fq.gz (86858983 sequences in total) Input files are E2_T1_C1_100_R1.fq.gz_C_to_T.fastq and E2_T1_C1_100_R1.fq.gz_G_to_A.fastq and E2_T1_C1_100_R2.fq.gz_C_to_T.fastq and E2_T1_C1_100_R2.fq.gz_G_to_A.fastq (FastQ) Now running 4 individual instances of Bowtie 2 against the bisulfite genome of /mmfs1/gscratch/scrubbed/sr320/github/Claudia-Halibut/project/data/genome/ with the specified options: -q --score-min L,0,-0.8 -p 8 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 Now starting a Bowtie 2 paired-end alignment for CTread1GAread2CTgenome (reading in sequences from E2_T1_C1_100_R1.fq.gz_C_to_T.fastq and E2_T1_C1_100_R2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.8 -p 8 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: lh00134:396:22JL32LT3:2:1101:1018:1064_1:N:0:GGACTTGG+NGTCTGCG:GGCAGTACG/1 77 * 0 0 * * 0 0 TNAATATAATTTTATAAAATTATAATAATATTATAATTTTTTATATATAAAATTAAGATTGGAAGAGTATATGTTTGAATTTTAGTTATGGATTTGGGGTAGTATGATTTGGGGGGGGGGGGGGGGGGGGGGGTTGGGGGGGGGGGGGGG -#IIIIIIIIIIIIIII-IIIIII9IIIIII9IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII-IIIIII9IIIIIIIIIIIIIIII9III-I-99IIIII9IIIII-I99I---99--II9IIIIIIIIII9 YT:Z:UP lh00134:396:22JL32LT3:2:1101:1018:1064_3:N:0:GGACTTGG+NGTCTGCG:GGCAGTACG/2 141 * 0 0 * * 0 0 TAATTTTATATATAAAAAATTATAATATTATTATAATTTTATAAAATTATATTTAAAATCAAAAAAACATCATATAAAAAAAAAATATCATCTACAATATAAATCTCAATAATCACCATATCACTAAAAAAAAAAAAAAAAAAAAAAAAA 9-I9IIIIIII9III99IIIII9IIIIIIIIIIIIIIIII9IIIIIIIIIIIIII9IIIIII9I9III9IIIIII-I-IIII9I9III-9I9--I-II-9-999----9I-99-I-I----99---999---9-999--II999-99999 YT:Z:UP Now starting a Bowtie 2 paired-end alignment for GAread1CTread2GAgenome (reading in sequences from E2_T1_C1_100_R1.fq.gz_G_to_A.fastq and E2_T1_C1_100_R2.fq.gz_C_to_T.fastq, with the options: -q --score-min L,0,-0.8 -p 8 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --norc)) Found first alignment: lh00134:396:22JL32LT3:2:1101:1018:1064_1:N:0:GGACTTGG+NGTCTGCG:GGCAGTACG/1 77 * 0 0 * * 0 0 CNAACACAACTCCATAAAACCACAACAACACTACAACCTTCTACACATAAAACCAAAATCAAAAAAACACACATCTAAACTCCAATCACAAACTTAAAACAATACAATCTAAAAAAAAAAAAAAAAAAAAAAATTAAAAAAAAAAAAAAA -#IIIIIIIIIIIIIII-IIIIII9IIIIII9IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII-IIIIII9IIIIIIIIIIIIIIII9III-I-99IIIII9IIIII-I99I---99--II9IIIIIIIIII9 YT:Z:UP lh00134:396:22JL32LT3:2:1101:1018:1064_3:N:0:GGACTTGG+NGTCTGCG:GGCAGTACG/2 141 * 0 0 * * 0 0 TGGTTTTATGTGTAGAAGGTTGTAGTGTTGTTGTGGTTTTATGGAGTTGTGTTTGAGATTGGAAGAGTGTTGTGTAGGGAAAGAGTGTTGTTTGTGGTGTGGATTTTGGTGGTTGTTGTGTTATTAAAAAGAGGGGGGGGGGGGGGGGGG 9-I9IIIIIII9III99IIIII9IIIIIIIIIIIIIIIII9IIIIIIIIIIIIII9IIIIII9I9III9IIIIII-I-IIII9I9III-9I9--I-II-9-999----9I-99-I-I----99---999---9-999--II999-99999 YT:Z:UP Now starting a Bowtie 2 paired-end alignment for GAread1CTread2CTgenome (reading in sequences from E2_T1_C1_100_R1.fq.gz_G_to_A.fastq and E2_T1_C1_100_R2.fq.gz_C_to_T.fastq, with the options: -q --score-min L,0,-0.8 -p 8 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --nofw)) Found first alignment: lh00134:396:22JL32LT3:2:1101:1018:1064_1:N:0:GGACTTGG+NGTCTGCG:GGCAGTACG/1 77 * 0 0 * * 0 0 CNAACACAACTCCATAAAACCACAACAACACTACAACCTTCTACACATAAAACCAAAATCAAAAAAACACACATCTAAACTCCAATCACAAACTTAAAACAATACAATCTAAAAAAAAAAAAAAAAAAAAAAATTAAAAAAAAAAAAAAA -#IIIIIIIIIIIIIII-IIIIII9IIIIII9IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII-IIIIII9IIIIIIIIIIIIIIII9III-I-99IIIII9IIIII-I99I---99--II9IIIIIIIIII9 YT:Z:UP lh00134:396:22JL32LT3:2:1101:1018:1064_3:N:0:GGACTTGG+NGTCTGCG:GGCAGTACG/2 141 * 0 0 * * 0 0 TGGTTTTATGTGTAGAAGGTTGTAGTGTTGTTGTGGTTTTATGGAGTTGTGTTTGAGATTGGAAGAGTGTTGTGTAGGGAAAGAGTGTTGTTTGTGGTGTGGATTTTGGTGGTTGTTGTGTTATTAAAAAGAGGGGGGGGGGGGGGGGGG 9-I9IIIIIII9III99IIIII9IIIIIIIIIIIIIIIII9IIIIIIIIIIIIII9IIIIII9I9III9IIIIII-I-IIII9I9III-9I9--I-II-9-999----9I-99-I-I----99---999---9-999--II999-99999 YT:Z:UP Now starting a Bowtie 2 paired-end alignment for CTread1GAread2GAgenome (reading in sequences from E2_T1_C1_100_R1.fq.gz_C_to_T.fastq and E2_T1_C1_100_R2.fq.gz_G_to_A.fastq, with the options: -q --score-min L,0,-0.8 -p 8 --reorder --ignore-quals --no-mixed --no-discordant --dovetail --maxins 500 --nofw)) Found first alignment: lh00134:396:22JL32LT3:2:1101:1018:1064_1:N:0:GGACTTGG+NGTCTGCG:GGCAGTACG/1 77 * 0 0 * * 0 0 TNAATATAATTTTATAAAATTATAATAATATTATAATTTTTTATATATAAAATTAAGATTGGAAGAGTATATGTTTGAATTTTAGTTATGGATTTGGGGTAGTATGATTTGGGGGGGGGGGGGGGGGGGGGGGTTGGGGGGGGGGGGGGG -#IIIIIIIIIIIIIII-IIIIII9IIIIII9IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII-IIIIII9IIIIIIIIIIIIIIII9III-I-99IIIII9IIIII-I99I---99--II9IIIIIIIIII9 YT:Z:UP lh00134:396:22JL32LT3:2:1101:1018:1064_3:N:0:GGACTTGG+NGTCTGCG:GGCAGTACG/2 141 * 0 0 * * 0 0 TAATTTTATATATAAAAAATTATAATATTATTATAATTTTATAAAATTATATTTAAAATCAAAAAAACATCATATAAAAAAAAAATATCATCTACAATATAAATCTCAATAATCACCATATCACTAAAAAAAAAAAAAAAAAAAAAAAAA 9-I9IIIIIII9III99IIIII9IIIIIIIIIIIIIIIII9IIIIIIIIIIIIII9IIIIII9I9III9IIIIII-I-IIII9I9III-9I9--I-II-9-999----9I-99-I-I----99---999---9-999--II999-99999 YT:Z:UP >>> Writing bisulfite mapping results to E2_T1_C1_100_pe.bam <<< Reading in the sequence files /mmfs1/gscratch/scrubbed/sr320/RRBS-run2/raw-data/E2_T1_C1_100_R1.fq.gz and /mmfs1/gscratch/scrubbed/sr320/RRBS-run2/raw-data/E2_T1_C1_100_R2.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 (ERR): bowtie2-align exited with value 141 (ERR): bowtie2-align exited with value 141 (ERR): bowtie2-align exited with value 141 (ERR): bowtie2-align exited with value 141