*** Bismark methylation extractor version v0.25.1 *** Output will be written into the directory: /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/ Summarising Bismark methylation extractor parameters: =============================================================== Bismark single-end SAM format specified (default) Number of cores to be used: 8 First 10 bp will be disregarded when processing the methylation call string Last 2 bp will be disregarded when processing the methylation call string Strand-specific outputs will be skipped. Separate output files for cytosines in CpG, CHG and CHH context will be generated Merge CHG and CHH context to non-CpG context specified Output path specified as: /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/ 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: '20G'. Temporary directory used for sorting is the output directory Checking file >>../data/methylation/dedup/SRR6995998.bam<< for signs of file truncation... Writing result file containing methylation information for C in CpG context to /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/CpG_context_SRR6995998.txt.gz Writing result file containing methylation information for C in any other context to /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/Non_CpG_context_SRR6995998.txt.gz Now reading in Bismark result file ../data/methylation/dedup/SRR6995998.bam Now reading in Bismark result file ../data/methylation/dedup/SRR6995998.bam Now reading in Bismark result file ../data/methylation/dedup/SRR6995998.bam Now reading in Bismark result file ../data/methylation/dedup/SRR6995998.bam Now reading in Bismark result file ../data/methylation/dedup/SRR6995998.bam Now reading in Bismark result file ../data/methylation/dedup/SRR6995998.bam Now reading in Bismark result file ../data/methylation/dedup/SRR6995998.bam Now reading in Bismark result file ../data/methylation/dedup/SRR6995998.bam skipping SAM header line: @HD VN:1.0 SO:unsorted skipping SAM header line: @SQ SN:NC_035780.1 LN:65668440 skipping SAM header line: @SQ SN:NC_035781.1 LN:61752955 skipping SAM header line: @SQ SN:NC_035782.1 LN:77061148 skipping SAM header line: @SQ SN:NC_035783.1 LN:59691872 skipping SAM header line: @SQ SN:NC_035784.1 LN:98698416 skipping SAM header line: @SQ SN:NC_035785.1 LN:51258098 skipping SAM header line: @SQ SN:NC_035786.1 LN:57830854 skipping SAM header line: @SQ SN:NC_035787.1 LN:75944018 skipping SAM header line: @SQ SN:NC_035788.1 LN:104168038 skipping SAM header line: @SQ SN:NC_035789.1 LN:32650045 skipping SAM header line: @SQ SN:NC_007175.2 LN:17244 skipping SAM header line: @PG ID:Bismark VN:v0.25.1 CL:"bismark --genome ../data/genome/bismark --non_directional --score_min L,0,-0.6 --parallel 6 --output_dir ../data/bismark --temp_dir ../data/bismark ../data/trimmed-reads/SRR6995998_trimmed.fq.gz" skipping SAM header line: @PG ID:samtools PN:samtools PP:Bismark VN:1.22.1 CL:/mmfs1/gscratch/srlab/sr320/miniforge3/envs/myflow/bin/samtools view -bSh - skipping SAM header line: @PG ID:samtools.1 PN:samtools PP:samtools VN:1.22.1 CL:/mmfs1/gscratch/srlab/sr320/miniforge3/envs/myflow/bin/samtools view -h /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/bismark/SRR6995998_trimmed.fq.gz.temp.1_bismark_bt2.bam skipping SAM header line: @PG ID:samtools.2 PN:samtools PP:samtools.1 VN:1.22.1 CL:/mmfs1/gscratch/srlab/sr320/miniforge3/envs/myflow/bin/samtools view --threads 4 -bSh - skipping SAM header line: @PG ID:samtools.3 PN:samtools PP:samtools.2 VN:1.22.1 CL:samtools view -h ../data/bismark/SRR6995998.bam skipping SAM header line: @PG ID:samtools.4 PN:samtools PP:samtools.3 VN:1.22.1 CL:samtools view -bS - skipping SAM header line: @PG ID:samtools.5 PN:samtools PP:samtools.4 VN:1.22.1 CL:/mmfs1/gscratch/srlab/sr320/miniforge3/envs/myflow/bin/samtools view -h --threads 1 ../data/bismark/SRR6995998.nonCG_filtered.bam skipping SAM header line: @PG ID:samtools.6 PN:samtools PP:samtools.5 VN:1.22.1 CL:/mmfs1/gscratch/srlab/sr320/miniforge3/envs/myflow/bin/samtools view -bS --threads 1 - skipping SAM header line: @PG ID:samtools.7 PN:samtools PP:samtools.6 VN:1.22.1 CL:/mmfs1/gscratch/srlab/sr320/miniforge3/envs/myflow/bin/samtools view -h ../data/methylation/dedup/SRR6995998.bam 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: 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: 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: 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 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 4000000 Processed lines: 4000000 Processed lines: 4000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 4000000 Processed lines: 4000000 Finished processing child process. Exiting.. Now waiting for all child processes to complete All child process successfully finished. Merging individual splitting reports into overall report: '/mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt' Merging from these individual files: /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.1 /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.2 /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.3 /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.4 /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.5 /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.6 /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.7 /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.8 Processed 4019827 lines in total Total number of methylation call strings processed: 4019827 Final Cytosine Methylation Report ================================= Total number of C's analysed: 65616268 Total methylated C's in CpG context: 4140274 Total methylated C's in CHG context: 182459 Total methylated C's in CHH context: 360704 Total C to T conversions in CpG context: 8394261 Total C to T conversions in CHG context: 15842074 Total C to T conversions in CHH context: 36696496 C methylated in CpG context: 33.0% C methylated in non-CpG context: 1.0% Merging individual M-bias reports into overall M-bias statistics from these 8 individual files: /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.1.mbias /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.2.mbias /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.3.mbias /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.4.mbias /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.5.mbias /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.6.mbias /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.7.mbias /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/SRR6995998_splitting_report.txt.8.mbias Determining maximum read length for M-Bias plot Maximum read length of Read 1: 89 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read length for M-Bias plot Maximum read length of Read 1: 89 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/CpG_context_SRR6995998.txt.gz contains data -> kept gzip: stdout: Broken pipe /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/Non_CpG_context_SRR6995998.txt.gz contains data -> kept Using these input files: /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/CpG_context_SRR6995998.txt.gz /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/Non_CpG_context_SRR6995998.txt.gz Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: SRR6995998.bedGraph.gz output directory: >/mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/< 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: 20G Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/CpG_context_SRR6995998.txt.gz Writing bedGraph to file: SRR6995998.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: SRR6995998.bismark.cov.gz Changed directory to /mmfs1/gscratch/scrubbed/sr320/github/project-gulf/data/methylation/dedup/ Now writing methylation information for file >>CpG_context_SRR6995998.txt.gz<< to individual files for each chromosome Finished writing out individual chromosome files for CpG_context_SRR6995998.txt.gz Collecting temporary chromosome file information... Processing the following input file(s): CpG_context_SRR6995998.txt.gz.chrNC_035787.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_035782.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_035781.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_035789.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_035788.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_035780.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_035784.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_035785.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_035783.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_035786.1.methXtractor.temp CpG_context_SRR6995998.txt.gz.chrNC_007175.2.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_007175.2.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_007175.2.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035780.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035780.1.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035781.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035781.1.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035782.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035782.1.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035783.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035783.1.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035784.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035784.1.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035785.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035785.1.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035786.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035786.1.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035787.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035787.1.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035788.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035788.1.methXtractor.temp Sorting input file CpG_context_SRR6995998.txt.gz.chrNC_035789.1.methXtractor.temp by positions (using -S of 20G) Successfully deleted the temporary input file CpG_context_SRR6995998.txt.gz.chrNC_035789.1.methXtractor.temp Finished BedGraph conversion ... gzip: stdout: Broken pipe