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/24B_1.fastq.gz /mmfs1/gscratch/coenv/sr320/cod-bs/24B_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/24B_1.fastq.gz and /mmfs1/gscratch/coenv/sr320/cod-bs/24B_2.fastq.gz Input files are in FastQ format Writing a C -> T converted version of the input file 24B_1.fastq.gz to 24B_1.fastq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file 24B_1.fastq.gz (64499170 sequences in total) Writing a G -> A converted version of the input file 24B_2.fastq.gz to 24B_2.fastq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file 24B_2.fastq.gz (64499170 sequences in total) Input files are 24B_1.fastq.gz_C_to_T.fastq and 24B_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 24B_1.fastq.gz_C_to_T.fastq and 24B_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: LH00160:538:22WG5VLT4:6:1101:23812:1042_1:N:0:GCTATCCT+AANAGGTG/1 99 NC_082388.1_CT_converted 8694974 4 133M8I10M = 8694966 -151 ANTAGTATTTTGTTAATGAGGGAATTGAAGTTATTTTTGTTTATGGTTGTTGGGTTATAGTTGGTTTTTTGTGGTTTTATTTTTTTTTTTTGATTGGTTTTGGTTAGATGTTTTTTGAGGGTTGGATGTAGTGGATTTTTAGATTGGAAGA I#IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII9IIIIIIIIIIII-IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII AS:i:-48 XS:i:-106 XN:i:0 XM:i:4 XO:i:1 XG:i:8 NM:i:12 MD:Z:1T136T2T0G0 YS:i:-38 YT:Z:CP LH00160:538:22WG5VLT4:6:1101:23812:1042_2:N:0:GCTATCCT+AANAGGTG/2 147 NC_082388.1_CT_converted 8694966 4 5M3I143M = 8694974 151 TTTTTTGATTTATTAGTATTTTGTTAATGAGGGAATTGAAGTTATTTTTGGTTATGGTTGTTGGGTTATAGTTGGTTTTTTGTGGTTTTATTTTTTTTTTTTGATTGGTTTTGGTTAGATGTTTTTTGAGGGTTGGATGTAGTGGATTTTT IIIIIIII9I9I9IIIIII9IIII9IIIIIIIIIIIIIIIIIII9III9I-II9I9IIIIIIII-IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII9IIIIIIIIIIIIIIIIII-IIIIIIIIIIIIIIIIIIIIIIIIIIIIIII AS:i:-38 XS:i:-97 XN:i:0 XM:i:4 XO:i:1 XG:i:3 NM:i:7 MD:Z:1A45T97G1A0 YS:i:-48 YT:Z:CP Now starting a Bowtie 2 paired-end alignment for CTread1GAread2GAgenome (reading in sequences from 24B_1.fastq.gz_C_to_T.fastq and 24B_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: LH00160:538:22WG5VLT4:6:1101:23812:1042_1:N:0:GCTATCCT+AANAGGTG/1 77 * 0 0 * * 0 0 ANTAGTATTTTGTTAATGAGGGAATTGAAGTTATTTTTGTTTATGGTTGTTGGGTTATAGTTGGTTTTTTGTGGTTTTATTTTTTTTTTTTGATTGGTTTTGGTTAGATGTTTTTTGAGGGTTGGATGTAGTGGATTTTTAGATTGGAAGA I#IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII9IIIIIIIIIIII-IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII YT:Z:UP LH00160:538:22WG5VLT4:6:1101:23812:1042_2:N:0:GCTATCCT+AANAGGTG/2 141 * 0 0 * * 0 0 AAAAATCCACTACATCCAACCCTCAAAAAACATCTAACCAAAACCAATCAAAAAAAAAAAATAAAACCACAAAAAACCAACTATAACCCAACAACCATAACCAAAAATAACTTCAATTCCCTCATTAACAAAATACTAATAAATCAAAAAA IIIIIIIIIIIIIIIIIIIIIIIIIIIIIII-IIIIIIIIIIIIIIIIII9IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII-IIIIIIII9I9II-I9III9IIIIIIIIIIIIIIIIIII9IIII9IIIIII9I9I9IIIIIIII YT:Z:UP >>> Writing bisulfite mapping results to 24B_pe.bam <<< Reading in the sequence files /mmfs1/gscratch/coenv/sr320/cod-bs/24B_1.fastq.gz and /mmfs1/gscratch/coenv/sr320/cod-bs/24B_2.fastq.gz Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1107:33782:18832_1:N:0:GCTATCCT+AACAGGTG NC_082399.1 21077814 Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1112:25787:13313_1:N:0:GCTATCCT+AACAGGTG NC_082386.1 22175832 Processed 1000000 sequence pairs so far Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1114:3549:2359_1:N:0:GCTATCCT+AACAGGTG NC_036931.1 16454 Processed 2000000 sequence pairs so far Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1130:34389:22348_1:N:0:GCTATCCT+AACAGGTG NC_082390.1 23393727 Processed 3000000 sequence pairs so far Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1139:7085:10805_1:N:0:GCTATCCT+AACAGGTG NC_082398.1 1 Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1145:11730:20583_1:N:0:GCTATCCT+AACAGGTG NC_036931.1 1 Processed 4000000 sequence pairs so far Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1154:30553:16577_1:N:0:GCTATCCT+AACAGGTG NC_082399.1 21077818 Processed 5000000 sequence pairs so far