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/63B_1.fastq.gz /mmfs1/gscratch/coenv/sr320/cod-bs/63B_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/63B_1.fastq.gz and /mmfs1/gscratch/coenv/sr320/cod-bs/63B_2.fastq.gz Input files are in FastQ format Writing a C -> T converted version of the input file 63B_1.fastq.gz to 63B_1.fastq.gz_C_to_T.fastq Created C -> T converted version of the FastQ file 63B_1.fastq.gz (59843638 sequences in total) Writing a G -> A converted version of the input file 63B_2.fastq.gz to 63B_2.fastq.gz_G_to_A.fastq Created G -> A converted version of the FastQ file 63B_2.fastq.gz (59843638 sequences in total) Input files are 63B_1.fastq.gz_C_to_T.fastq and 63B_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 63B_1.fastq.gz_C_to_T.fastq and 63B_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: LH00160:538:22WG5VLT4:6:1101:23051:1042_1:N:0:CCTTGATC+GANGGAGT/1 99 NC_082397.1_CT_converted 27110892 1 151M = 27110953 212 ANATTGTTGATTTTATTTAAAATGAATGTATTTTATTTTTAAATGGTTTAGTATGGTTTAATAATTTTTTAGTAAGTTTAGTAATATGTTTGAATTGAAATTGATATAGATATTAATGTTTTTTGTTTTTATGTTTGTAAAATGTTTTTTA I#IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII9IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII AS:i:-1 XS:i:-1 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:1T149 YS:i:-6 YT:Z:CP LH00160:538:22WG5VLT4:6:1101:23051:1042_2:N:0:CCTTGATC+GANGGAGT/2 147 NC_082397.1_CT_converted 27110953 1 151M = 27110892 -212 TAATTTTTTAGTAAGTTTAGTAATATGTTTGAATTGAAATTGATATAGATATTAATGTTTTTTGTTTTTATGTTTGTAAAATGTTTTTTAGAGTGTTAGAATTATTTTTTGATAGATAATGTGTTTTTGATTTTATATGGTTGATTTTTTT IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII-IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII AS:i:-6 XS:i:-6 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:148A2 YS:i:-1 YT:Z:CP Now starting a Bowtie 2 paired-end alignment for CTread1GAread2GAgenome (reading in sequences from 63B_1.fastq.gz_C_to_T.fastq and 63B_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: LH00160:538:22WG5VLT4:6:1101:23051:1042_1:N:0:CCTTGATC+GANGGAGT/1 83 NC_082401.1_GA_converted 3308707 1 151M = 3308646 -212 TAAAAAACATTTTACAAACATAAAAACAAAAAACATTAATATCTATATCAATTTCAATTCAAACATATTACTAAACTTACTAAAAAATTATTAAACCATACTAAACCATTTAAAAATAAAATACATTCATTTTAAATAAAATCAACAATNT IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII9IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII#I AS:i:-1 XS:i:-1 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:149A1 YS:i:-6 YT:Z:CP LH00160:538:22WG5VLT4:6:1101:23051:1042_2:N:0:CCTTGATC+GANGGAGT/2 163 NC_082401.1_GA_converted 3308646 1 151M = 3308707 212 AAAAAAATCAACCATATAAAATCAAAAACACATTATCTATCAAAAAATAATTCTAACACTCTAAAAAACATTTTACAAACATAAAAACAAAAAACATTAATATCTATATCAATTTCAATTCAAACATATTACTAAACTTACTAAAAAATTA IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII-IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII AS:i:-6 XS:i:-6 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:2T148 YS:i:-1 YT:Z:CP >>> Writing bisulfite mapping results to 63B_pe.bam <<< Reading in the sequence files /mmfs1/gscratch/coenv/sr320/cod-bs/63B_1.fastq.gz and /mmfs1/gscratch/coenv/sr320/cod-bs/63B_2.fastq.gz Chromosomal sequence could not be extracted for LH00160:538:22WG5VLT4:6:1107:43614:27979_1:N:0:CCTTGATC+GATGGAGT NC_082382.1 26289603 Processed 1000000 sequence pairs so far