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/project-cod-temperature/data/genome/ (absolute path is '/mmfs1/gscratch/scrubbed/sr320/github/project-cod-temperature/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/project-cod-temperature/output/16-bismark'): /mmfs1/gscratch/coenv/sr320/cod-bs/02B_1.fastq.gz /mmfs1/gscratch/coenv/sr320/cod-bs/02B_2.fastq.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!) Output will be written into the directory: /mmfs1/gscratch/scrubbed/sr320/github/project-cod-temperature/output/16-bismark/ 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/project-cod-temperature/output/16-bismark Now reading in and storing sequence information of the genome specified in: /mmfs1/gscratch/scrubbed/sr320/github/project-cod-temperature/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/coenv/sr320/cod-bs/02B_1.fastq.gz and /mmfs1/gscratch/coenv/sr320/cod-bs/02B_2.fastq.gz Input files are in FastQ format Writing a C -> T converted version of the input file 02B_1.fastq.gz to 02B_1.fastq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file 02B_1.fastq.gz (54080874 sequences in total) Writing a G -> A converted version of the input file 02B_2.fastq.gz to 02B_2.fastq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file 02B_2.fastq.gz (54080874 sequences in total) Input files are 02B_1.fastq.gz_C_to_T.fastq and 02B_2.fastq.gz_G_to_A.fastq (FastQ) Now running 2 instances of Bowtie 2 against the bisulfite genome of /mmfs1/gscratch/scrubbed/sr320/github/project-cod-temperature/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 02B_1.fastq.gz_C_to_T.fastq and 02B_2.fastq.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: LH00160:538:22WG5VLT4:6:1101:22016:1042_1:N:0:ACTCTCGA+CTNTACCA/1 77 * 0 0 * * 0 0 TNATAGGATGTGGATAGTAGTTAAATTGAGAGATTGGGAATGTTAGTTATTTGGTTTATTGGATTTTGAATGTGATATTTGTTTGTTAAGGTTTTAGTAGGATAAGATAATGTTTTTTTTTAGATTGGAAGAGTATATGTTTGAATTTTAG I#IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII9IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII YT:Z:UP LH00160:538:22WG5VLT4:6:1101:22016:1042_2:N:0:ACTCTCGA+CTNTACCA/2 141 * 0 0 * * 0 0 AAAAAAAAACATTATCTTATCCTACTAAAACCTTAACAAACAAATATCACATTCAAAATCCAATAAACCAAATAACTAACATTCCCAATCTCTCAATTTAACTACTATCCACATCCTATAAAAATCAAAAAAACATCATATAAAAAAAAAA IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII9IIIIIIII99 YT:Z:UP Now starting a Bowtie 2 paired-end alignment for CTread1GAread2GAgenome (reading in sequences from 02B_1.fastq.gz_C_to_T.fastq and 02B_2.fastq.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: LH00160:538:22WG5VLT4:6:1101:22016:1042_1:N:0:ACTCTCGA+CTNTACCA/1 83 NC_082395.1_GA_converted 4585353 0 18M3I2M14I114M = 4585368 150 CTAAAATTCAAACATATACTCTTCCAATCTAAAAAAAAACATTATCTTATCCTACTAAAACCTTAACAAACAAATATCACATTCAAAATCCAATAAACCAAATAACTAACATTCCCAATCTCTCAATTTAACTACTATCCACATCCTATNA IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII9IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII#I AS:i:-98 XN:i:0 XM:i:7 XO:i:2 XG:i:17 NM:i:24 MD:Z:0A1T6C0T1T0T118A1 YS:i:-111 YT:Z:CP LH00160:538:22WG5VLT4:6:1101:22016:1042_2:N:0:ACTCTCGA+CTNTACCA/2 163 NC_082395.1_GA_converted 4585368 0 4M2I113M8I14M6I4M = 4585353 -150 AAAAAAAAACATTATCTTATCCTACTAAAACCTTAACAAACAAATATCACATTCAAAATCCAATAAACCAAATAACTAACATTCCCAATCTCTCAATTTAACTACTATCCACATCCTATAAAAATCAAAAAAACATCATATAAAAAAAAAA IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII9IIIIIIII99 AS:i:-111 XN:i:0 XM:i:8 XO:i:3 XG:i:16 NM:i:24 MD:Z:1T1T0T119C2T0C0C4T0 YS:i:-98 YT:Z:CP >>> Writing bisulfite mapping results to 02B_pe.bam <<< Reading in the sequence files /mmfs1/gscratch/coenv/sr320/cod-bs/02B_1.fastq.gz and /mmfs1/gscratch/coenv/sr320/cod-bs/02B_2.fastq.gz Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1104:6802:22362_1:N:0:ACTCTCGA+CTGTACCA NC_082396.1 24646082 Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1104:6794:22376_1:N:0:ACTCTCGA+CTGTACCA NC_082396.1 24646082 Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1113:19199:13733_1:N:0:ACTCTCGA+CTGTACCA NC_082398.1 1 Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1113:19216:13733_1:N:0:ACTCTCGA+CTGTACCA NC_082398.1 1 Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1113:19224:13747_1:N:0:ACTCTCGA+CTGTACCA NC_082398.1 1 Processed 1000000 sequence pairs so far Processed 2000000 sequence pairs so far Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1157:6705:3171_1:N:0:ACTCTCGA+CTGTACCA NC_082398.1 1 Processed 3000000 sequence pairs so far Processed 4000000 sequence pairs so far Processed 5000000 sequence pairs so far