FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 28 threads. Please monitor performance closely and tune down if needed! chr NC_035780.1 (65668440 bp) chr NC_035781.1 (61752955 bp) chr NC_035782.1 (77061148 bp) chr NC_035783.1 (59691872 bp) chr NC_035784.1 (98698416 bp) chr NC_035785.1 (51258098 bp) chr NC_035786.1 (57830854 bp) chr NC_035787.1 (75944018 bp) chr NC_035788.1 (104168038 bp) chr NC_035789.1 (32650045 bp) chr NC_007175.2 (17244 bp) Number of paired-end alignments with a unique best hit: 13794188 Mapping efficiency: 25.0% Sequence pairs with no alignments under any condition: 34816649 Sequence pairs did not map uniquely: 6572017 Sequence pairs which were discarded because genomic sequence could not be extracted: 1 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 3478061 ((converted) top strand) GA/CT/CT: 3412272 (complementary to (converted) top strand) GA/CT/GA: 3419707 (complementary to (converted) bottom strand) CT/GA/GA: 3484147 ((converted) bottom strand) *** Bismark methylation extractor version v0.19.0 *** Trying to determine the type of mapping from the SAM header line of file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Treating file(s) as paired-end data (as extracted from @PG line) Setting option '--no_overlap' since this is (normally) the right thing to do for paired-end data Core usage currently set to more than 20 threads. Let's see how this goes... (set value: 28) Summarising Bismark methylation extractor parameters: =============================================================== Bismark paired-end SAM format specified (default) Number of cores to be used: 28 Output will be written to the current directory ('/gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/avg_reads_bismark') 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) White spaces in read ID names will be removed prior to sorting The bedGraph UNIX sort command will use the following memory setting: '75%'. Temporary directory used for sorting is the output directory Checking file >>cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.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:v0.19.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ --genome /gscratch/srlab/sam/data/C_virginica/genomes/GCF_002022765.2_C_virginica-3.0/ --non_directional --score_min L,0,-0.6 -u 55182854 -p 28 -1 /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/cvir_bsseq_all_pe_R1.fastq.gz -2 /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/cvir_bsseq_all_pe_R2.fastq.gz" 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: 500000 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: 500000 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: 500000 Processed lines: 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Exiting.. Processed lines: 13000000 Processed lines: 13500000 Processed lines: 13500000 Processed lines: 13500000 Processed lines: 13000000 Finished processing child process. Exiting.. Processed lines: 13500000 Processed lines: 13500000 Now waiting for all child processes to complete Processed lines: 13500000 Processed lines: 13500000 Finished processing child process. Exiting.. Processed lines: 13500000 Processed lines: 13500000 Finished processing child process. Exiting.. Processed lines: 13500000 Processed lines: 13500000 Processed lines: 13500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 13500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 13500000 Processed lines: 13500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 13500000 Finished processing child process. Exiting.. 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Merging individual splitting reports into overall report: 'cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt' Merging from these individual files: cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.1 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.2 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.3 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.4 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.5 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.6 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.7 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.8 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.9 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.10 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.11 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.12 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.13 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.14 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.15 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.16 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.17 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.18 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.19 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.20 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.21 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.22 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.23 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.24 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.25 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.26 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.27 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.28 Processed 13794187 lines in total Total number of methylation call strings processed: 27588374 Final Cytosine Methylation Report ================================= Total number of C's analysed: 244017867 Total methylated C's in CpG context: 24952246 Total methylated C's in CHG context: 1127439 Total methylated C's in CHH context: 4022627 Total C to T conversions in CpG context: 8868175 Total C to T conversions in CHG context: 53972893 Total C to T conversions in CHH context: 151074487 C methylated in CpG context: 73.8% C methylated in CHG context: 2.0% C methylated in CHH context: 2.6% Merging individual M-bias reports into overall M-bias statistics from these 28 individual files: cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.1.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.2.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.3.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.4.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.5.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.6.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.7.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.8.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.9.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.10.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.11.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.12.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.13.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.14.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.15.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.16.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.17.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.18.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.19.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.20.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.21.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.22.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.23.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.24.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.25.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.26.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.27.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.28.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 80 Maximum read length of Read 2: 80 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 80 Maximum read length of Read 2: 80 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept Using these input files: CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz output directory: >< remove whitespaces: yes CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/avg_reads_bismark/CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/avg_reads_bismark/CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/avg_reads_bismark/CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/avg_reads_bismark/CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing bedGraph to file: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/avg_reads_bismark/CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/avg_reads_bismark/CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/avg_reads_bismark/CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/avg_reads_bismark/CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Sorting input file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Found 1 alignment reports in current directory. Now trying to figure out whether there are corresponding optional reports Writing Bismark HTML report to >> cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt < Processing alignment report cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt ... Complete No deduplication report file specified, skipping this step Using the following splitting report: > cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt < Processing splitting report cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt ... Complete Using the following M-bias report: > cvir_bsseq_all_pe_R1_bismark_bt2_pe.M-bias.txt < Processing M-bias report cvir_bsseq_all_pe_R1_bismark_bt2_pe.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== No Bismark/Bowtie2 single-end BAM files detected Found Bismark/Bowtie2 paired-end files No Bismark/Bowtie single-end BAM files detected No Bismark/Bowtie paired-end BAM files detected Generating Bismark summary report from 1 Bismark BAM file(s)... >> Reading from Bismark report: cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt No deduplication report present, skipping... No methylation extractor report present, skipping... Wrote Bismark project summary to >> bismark_summary_report.html << [bam_sort_core] merging from 0 files and 28 in-memory blocks... FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 28 threads. Please monitor performance closely and tune down if needed! chr NC_035780.1 (65668440 bp) chr NC_035781.1 (61752955 bp) chr NC_035782.1 (77061148 bp) chr NC_035783.1 (59691872 bp) chr NC_035784.1 (98698416 bp) chr NC_035785.1 (51258098 bp) chr NC_035786.1 (57830854 bp) chr NC_035787.1 (75944018 bp) chr NC_035788.1 (104168038 bp) chr NC_035789.1 (32650045 bp) chr NC_007175.2 (17244 bp) Number of paired-end alignments with a unique best hit: 7964108 Mapping efficiency: 28.9% Sequence pairs with no alignments under any condition: 15945689 Sequence pairs did not map uniquely: 3681630 Sequence pairs which were discarded because genomic sequence could not be extracted: 0 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 2015219 ((converted) top strand) GA/CT/CT: 1962001 (complementary to (converted) top strand) GA/CT/GA: 1965430 (complementary to (converted) bottom strand) CT/GA/GA: 2021458 ((converted) bottom strand) *** Bismark methylation extractor version v0.19.0 *** Trying to determine the type of mapping from the SAM header line of file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Treating file(s) as paired-end data (as extracted from @PG line) Setting option '--no_overlap' since this is (normally) the right thing to do for paired-end data Core usage currently set to more than 20 threads. Let's see how this goes... (set value: 28) Summarising Bismark methylation extractor parameters: =============================================================== Bismark paired-end SAM format specified (default) Number of cores to be used: 28 Output will be written to the current directory ('/gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_avg_reads_bismark') 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) White spaces in read ID names will be removed prior to sorting The bedGraph UNIX sort command will use the following memory setting: '75%'. Temporary directory used for sorting is the output directory Checking file >>cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.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:v0.19.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ --genome /gscratch/srlab/sam/data/C_virginica/genomes/GCF_002022765.2_C_virginica-3.0/ --non_directional --score_min L,0,-0.6 -u 27591427 -p 28 -1 /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/cvir_bsseq_all_pe_R1.fastq.gz -2 /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/cvir_bsseq_all_pe_R2.fastq.gz" 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: 500000 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: 500000 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: 500000 Processed lines: 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Exiting.. Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Finished processing child process. Exiting.. Processed lines: 7500000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7000000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Finished processing child process. Exiting.. Processed lines: 7500000 Finished processing child process. Exiting.. Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Finished processing child process. Exiting.. Processed lines: 7500000 Finished processing child process. Exiting.. Processed lines: 7500000 Processed lines: 7500000 Processed lines: 7500000 Now waiting for all child processes to complete Processed lines: 7500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. 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Merging individual splitting reports into overall report: 'cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt' Merging from these individual files: cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.1 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.2 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.3 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.4 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.5 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.6 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.7 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.8 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.9 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.10 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.11 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.12 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.13 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.14 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.15 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.16 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.17 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.18 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.19 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.20 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.21 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.22 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.23 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.24 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.25 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.26 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.27 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.28 Processed 7964108 lines in total Total number of methylation call strings processed: 15928216 Final Cytosine Methylation Report ================================= Total number of C's analysed: 145927935 Total methylated C's in CpG context: 15041047 Total methylated C's in CHG context: 689452 Total methylated C's in CHH context: 2237441 Total C to T conversions in CpG context: 5505093 Total C to T conversions in CHG context: 32294077 Total C to T conversions in CHH context: 90160825 C methylated in CpG context: 73.2% C methylated in CHG context: 2.1% C methylated in CHH context: 2.4% Merging individual M-bias reports into overall M-bias statistics from these 28 individual files: cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.1.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.2.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.3.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.4.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.5.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.6.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.7.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.8.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.9.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.10.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.11.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.12.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.13.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.14.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.15.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.16.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.17.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.18.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.19.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.20.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.21.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.22.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.23.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.24.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.25.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.26.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.27.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.28.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 80 Maximum read length of Read 2: 80 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 80 Maximum read length of Read 2: 80 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept Using these input files: CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz output directory: >< remove whitespaces: yes CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_avg_reads_bismark/CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_avg_reads_bismark/CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_avg_reads_bismark/CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_avg_reads_bismark/CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing bedGraph to file: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_avg_reads_bismark/CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_avg_reads_bismark/CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_avg_reads_bismark/CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_avg_reads_bismark/CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Sorting input file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Found 1 alignment reports in current directory. Now trying to figure out whether there are corresponding optional reports Writing Bismark HTML report to >> cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt < Processing alignment report cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt ... Complete No deduplication report file specified, skipping this step Using the following splitting report: > cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt < Processing splitting report cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt ... Complete Using the following M-bias report: > cvir_bsseq_all_pe_R1_bismark_bt2_pe.M-bias.txt < Processing M-bias report cvir_bsseq_all_pe_R1_bismark_bt2_pe.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== No Bismark/Bowtie2 single-end BAM files detected Found Bismark/Bowtie2 paired-end files No Bismark/Bowtie single-end BAM files detected No Bismark/Bowtie paired-end BAM files detected Generating Bismark summary report from 1 Bismark BAM file(s)... >> Reading from Bismark report: cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt No deduplication report present, skipping... No methylation extractor report present, skipping... Wrote Bismark project summary to >> bismark_summary_report.html << [bam_sort_core] merging from 0 files and 28 in-memory blocks... FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 28 threads. Please monitor performance closely and tune down if needed! chr NC_035780.1 (65668440 bp) chr NC_035781.1 (61752955 bp) chr NC_035782.1 (77061148 bp) chr NC_035783.1 (59691872 bp) chr NC_035784.1 (98698416 bp) chr NC_035785.1 (51258098 bp) chr NC_035786.1 (57830854 bp) chr NC_035787.1 (75944018 bp) chr NC_035788.1 (104168038 bp) chr NC_035789.1 (32650045 bp) chr NC_007175.2 (17244 bp) Number of paired-end alignments with a unique best hit: 90400225 Mapping efficiency: 32.8% Sequence pairs with no alignments under any condition: 142295313 Sequence pairs did not map uniquely: 43218734 Sequence pairs which were discarded because genomic sequence could not be extracted: 9 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 23103068 ((converted) top strand) GA/CT/CT: 22071301 (complementary to (converted) top strand) GA/CT/GA: 22087710 (complementary to (converted) bottom strand) CT/GA/GA: 23138137 ((converted) bottom strand) *** Bismark methylation extractor version v0.19.0 *** Trying to determine the type of mapping from the SAM header line of file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Treating file(s) as paired-end data (as extracted from @PG line) Setting option '--no_overlap' since this is (normally) the right thing to do for paired-end data Core usage currently set to more than 20 threads. Let's see how this goes... (set value: 28) Summarising Bismark methylation extractor parameters: =============================================================== Bismark paired-end SAM format specified (default) Number of cores to be used: 28 Output will be written to the current directory ('/gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_total_reads_bismark') 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) White spaces in read ID names will be removed prior to sorting The bedGraph UNIX sort command will use the following memory setting: '75%'. Temporary directory used for sorting is the output directory Checking file >>cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.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:v0.19.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ --genome /gscratch/srlab/sam/data/C_virginica/genomes/GCF_002022765.2_C_virginica-3.0/ --non_directional --score_min L,0,-0.6 -u 275914272 -p 28 -1 /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/cvir_bsseq_all_pe_R1.fastq.gz -2 /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/cvir_bsseq_all_pe_R2.fastq.gz" 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: 500000 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: 500000 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: 500000 Processed lines: 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Exiting.. 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Exiting.. 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Processed lines: 89000000 Processed lines: 89000000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 89000000 Processed lines: 88000000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 89000000 Processed lines: 88000000 Processed lines: 89000000 Processed lines: 88000000 Now waiting for all child processes to complete Processed lines: 89000000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 89500000 Processed lines: 89500000 Processed lines: 89500000 Processed lines: 89500000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 90000000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 88500000 Processed lines: 89500000 Processed lines: 89500000 Processed lines: 88500000 Processed lines: 89500000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 89000000 Processed lines: 89000000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 89500000 Processed lines: 90000000 Processed lines: 90000000 Processed lines: 90000000 Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 89500000 Processed lines: 90000000 Processed lines: 89000000 Processed lines: 90000000 Processed lines: 89500000 Processed lines: 90000000 Processed lines: 89500000 Processed lines: 90000000 Processed lines: 90000000 Processed lines: 89000000 Processed lines: 90000000 Processed lines: 89000000 Processed lines: 90000000 Processed lines: 90000000 Processed lines: 89500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 89500000 Processed lines: 89500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 89500000 Processed lines: 90000000 Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 90000000 Processed lines: 89500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 89500000 Finished processing child process. Exiting.. Processed lines: 89500000 Processed lines: 90000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 90000000 Processed lines: 90000000 Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 90000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 90000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt' Merging from these individual files: cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.1 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.2 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.3 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.4 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.5 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.6 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.7 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.8 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.9 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.10 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.11 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.12 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.13 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.14 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.15 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.16 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.17 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.18 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.19 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.20 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.21 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.22 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.23 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.24 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.25 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.26 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.27 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.28 Processed 90400216 lines in total Total number of methylation call strings processed: 180800432 Final Cytosine Methylation Report ================================= Total number of C's analysed: 1564663196 Total methylated C's in CpG context: 159187724 Total methylated C's in CHG context: 7202288 Total methylated C's in CHH context: 24250798 Total C to T conversions in CpG context: 52805909 Total C to T conversions in CHG context: 346029728 Total C to T conversions in CHH context: 975186749 C methylated in CpG context: 75.1% C methylated in CHG context: 2.0% C methylated in CHH context: 2.4% Merging individual M-bias reports into overall M-bias statistics from these 28 individual files: cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.1.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.2.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.3.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.4.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.5.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.6.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.7.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.8.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.9.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.10.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.11.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.12.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.13.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.14.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.15.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.16.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.17.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.18.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.19.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.20.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.21.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.22.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.23.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.24.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.25.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.26.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.27.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.28.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 80 Maximum read length of Read 2: 80 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 80 Maximum read length of Read 2: 80 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept Using these input files: CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz output directory: >< remove whitespaces: yes CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_total_reads_bismark/CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_total_reads_bismark/CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_total_reads_bismark/CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_total_reads_bismark/CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing bedGraph to file: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_total_reads_bismark/CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_total_reads_bismark/CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_total_reads_bismark/CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/half_total_reads_bismark/CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Sorting input file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Found 1 alignment reports in current directory. Now trying to figure out whether there are corresponding optional reports Writing Bismark HTML report to >> cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt < Processing alignment report cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt ... Complete No deduplication report file specified, skipping this step Using the following splitting report: > cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt < Processing splitting report cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt ... Complete Using the following M-bias report: > cvir_bsseq_all_pe_R1_bismark_bt2_pe.M-bias.txt < Processing M-bias report cvir_bsseq_all_pe_R1_bismark_bt2_pe.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== No Bismark/Bowtie2 single-end BAM files detected Found Bismark/Bowtie2 paired-end files No Bismark/Bowtie single-end BAM files detected No Bismark/Bowtie paired-end BAM files detected Generating Bismark summary report from 1 Bismark BAM file(s)... >> Reading from Bismark report: cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt No deduplication report present, skipping... No methylation extractor report present, skipping... Wrote Bismark project summary to >> bismark_summary_report.html << [bam_sort_core] merging from 56 files and 28 in-memory blocks... FastQ format assumed (by default) Each Bowtie 2 instance is going to be run with 28 threads. Please monitor performance closely and tune down if needed! chr NC_035780.1 (65668440 bp) chr NC_035781.1 (61752955 bp) chr NC_035782.1 (77061148 bp) chr NC_035783.1 (59691872 bp) chr NC_035784.1 (98698416 bp) chr NC_035785.1 (51258098 bp) chr NC_035786.1 (57830854 bp) chr NC_035787.1 (75944018 bp) chr NC_035788.1 (104168038 bp) chr NC_035789.1 (32650045 bp) chr NC_007175.2 (17244 bp) Number of paired-end alignments with a unique best hit: 90400225 Mapping efficiency: 32.8% Sequence pairs with no alignments under any condition: 142295313 Sequence pairs did not map uniquely: 43218734 Sequence pairs which were discarded because genomic sequence could not be extracted: 9 Number of sequence pairs with unique best (first) alignment came from the bowtie output: CT/GA/CT: 23103068 ((converted) top strand) GA/CT/CT: 22071301 (complementary to (converted) top strand) GA/CT/GA: 22087710 (complementary to (converted) bottom strand) CT/GA/GA: 23138137 ((converted) bottom strand) *** Bismark methylation extractor version v0.19.0 *** Trying to determine the type of mapping from the SAM header line of file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Treating file(s) as paired-end data (as extracted from @PG line) Setting option '--no_overlap' since this is (normally) the right thing to do for paired-end data Core usage currently set to more than 20 threads. Let's see how this goes... (set value: 28) Summarising Bismark methylation extractor parameters: =============================================================== Bismark paired-end SAM format specified (default) Number of cores to be used: 28 Output will be written to the current directory ('/gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/total_reads_bismark') 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) White spaces in read ID names will be removed prior to sorting The bedGraph UNIX sort command will use the following memory setting: '75%'. Temporary directory used for sorting is the output directory Checking file >>cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam<< for signs of file truncation... Now testing Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Writing result file containing methylation information for C in CpG context from the original top strand to CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the complementary to original top strand to CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the complementary to original bottom strand to CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CpG context from the original bottom strand to CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the original top strand to CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the complementary to original top strand to CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the complementary to original bottom strand to CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHG context from the original bottom strand to CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the original top strand to CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the complementary to original top strand to CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the complementary to original bottom strand to CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing result file containing methylation information for C in CHH context from the original bottom strand to CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bam Now reading in Bismark result file cvir_bsseq_all_pe_R1_bismark_bt2_pe.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:v0.19.0 CL:"bismark --path_to_bowtie /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ --genome /gscratch/srlab/sam/data/C_virginica/genomes/GCF_002022765.2_C_virginica-3.0/ --non_directional --score_min L,0,-0.6 -p 28 -1 /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/cvir_bsseq_all_pe_R1.fastq.gz -2 /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/cvir_bsseq_all_pe_R2.fastq.gz" 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: 500000 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: 500000 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: 500000 Processed lines: 500000 Processed 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Exiting.. 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88000000 Processed lines: 89000000 Processed lines: 88000000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 88000000 Processed lines: 88000000 Processed lines: 90000000 Processed lines: 88500000 Processed lines: 89500000 Processed lines: 88000000 Processed lines: 88500000 Processed lines: 88000000 Processed lines: 88500000 Processed lines: 88500000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 88000000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 88500000 Processed lines: 89500000 Processed lines: 88500000 Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 89500000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 88500000 Processed lines: 90000000 Processed lines: 89000000 Processed lines: 88500000 Processed lines: 89000000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 90000000 Processed lines: 90000000 Processed lines: 88500000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 89000000 Now waiting for all child processes to complete Processed lines: 90000000 Processed lines: 89000000 Finished processing child process. Exiting.. Processed lines: 90000000 Processed lines: 90000000 Processed lines: 89500000 Processed lines: 90000000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 89000000 Processed lines: 89500000 Processed lines: 89000000 Processed lines: 89500000 Finished processing child process. Exiting.. Processed lines: 89500000 Finished processing child process. Exiting.. Processed lines: 89000000 Processed lines: 89500000 Processed lines: 90000000 Processed lines: 89000000 Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 89500000 Processed lines: 89500000 Processed lines: 90000000 Processed lines: 89500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 89500000 Finished processing child process. Exiting.. Processed lines: 90000000 Processed lines: 90000000 Processed lines: 89500000 Processed lines: 89500000 Processed lines: 90000000 Processed lines: 89500000 Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 90000000 Processed lines: 90000000 Processed lines: 89500000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 89500000 Processed lines: 90000000 Processed lines: 90000000 Processed lines: 90000000 Processed lines: 90000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 90000000 Processed lines: 90000000 Finished processing child process. Exiting.. Processed lines: 90000000 Processed lines: 90000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Processed lines: 90000000 Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Finished processing child process. Exiting.. Merging individual splitting reports into overall report: 'cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt' Merging from these individual files: cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.1 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.2 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.3 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.4 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.5 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.6 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.7 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.8 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.9 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.10 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.11 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.12 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.13 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.14 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.15 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.16 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.17 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.18 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.19 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.20 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.21 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.22 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.23 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.24 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.25 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.26 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.27 cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.28 Processed 90400216 lines in total Total number of methylation call strings processed: 180800432 Final Cytosine Methylation Report ================================= Total number of C's analysed: 1564663196 Total methylated C's in CpG context: 159187724 Total methylated C's in CHG context: 7202288 Total methylated C's in CHH context: 24250798 Total C to T conversions in CpG context: 52805909 Total C to T conversions in CHG context: 346029728 Total C to T conversions in CHH context: 975186749 C methylated in CpG context: 75.1% C methylated in CHG context: 2.0% C methylated in CHH context: 2.4% Merging individual M-bias reports into overall M-bias statistics from these 28 individual files: cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.1.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.2.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.3.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.4.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.5.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.6.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.7.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.8.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.9.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.10.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.11.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.12.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.13.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.14.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.15.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.16.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.17.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.18.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.19.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.20.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.21.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.22.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.23.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.24.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.25.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.26.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.27.mbias cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt.28.mbias Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 80 Maximum read length of Read 2: 80 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Determining maximum read lengths for M-Bias plots Maximum read length of Read 1: 80 Maximum read length of Read 2: 80 Perl module GD::Graph::lines is not installed, skipping drawing M-bias plots (only writing out M-bias plot table) Deleting unused files ... CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt contains data -> kept Using these input files: CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt CHH_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Summary of parameters for bismark2bedGraph conversion: ====================================================== bedGraph output: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz output directory: >< remove whitespaces: yes CX context: no (CpG context only, default) No-header selected: no Sorting method: Unix sort-based (smaller memory footprint, but slower) Sort buffer size: 75% Coverage threshold: 1 ============================================================================= Methylation information will now be written into a bedGraph and coverage file ============================================================================= Using the following files as Input: /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/total_reads_bismark/CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/total_reads_bismark/CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/total_reads_bismark/CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/total_reads_bismark/CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt Writing bedGraph to file: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz Also writing out a coverage file including counts methylated and unmethylated residues to file: cvir_bsseq_all_pe_R1_bismark_bt2_pe.bismark.cov.gz The genome of interest was specified to contain gazillions of chromosomes or scaffolds. Merging all input files and sorting everything in memory instead of writing out individual chromosome files... Writing all merged methylation calls to temp file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/total_reads_bismark/CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_OT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/total_reads_bismark/CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_CTOT_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/total_reads_bismark/CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_CTOB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Now replacing whitespaces in the sequence ID field of the Bismark methylation extractor output /gscratch/scrubbed/samwhite/outputs/20190312_cvir_gonad_bismark/total_reads_bismark/CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt prior to bedGraph conversion Attempting to write to file CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt Finished writing methylation calls from CpG_OB_cvir_bsseq_all_pe_R1_bismark_bt2_pe.txt.spaces_removed.txt to merged temp file Sorting input file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged by positions (using -S of 75%) Successfully deleted the temporary input file cvir_bsseq_all_pe_R1_bismark_bt2_pe.bedGraph.gz.methylation_calls.merged Finished BedGraph conversion ... Found 1 alignment reports in current directory. Now trying to figure out whether there are corresponding optional reports Writing Bismark HTML report to >> cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.html << ============================================================================================================== Using the following alignment report: > cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt < Processing alignment report cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt ... Complete No deduplication report file specified, skipping this step Using the following splitting report: > cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt < Processing splitting report cvir_bsseq_all_pe_R1_bismark_bt2_pe_splitting_report.txt ... Complete Using the following M-bias report: > cvir_bsseq_all_pe_R1_bismark_bt2_pe.M-bias.txt < Processing M-bias report cvir_bsseq_all_pe_R1_bismark_bt2_pe.M-bias.txt ... Complete No nucleotide coverage report file specified, skipping this step ============================================================================================================== No Bismark/Bowtie2 single-end BAM files detected Found Bismark/Bowtie2 paired-end files No Bismark/Bowtie single-end BAM files detected No Bismark/Bowtie paired-end BAM files detected Generating Bismark summary report from 1 Bismark BAM file(s)... >> Reading from Bismark report: cvir_bsseq_all_pe_R1_bismark_bt2_PE_report.txt No deduplication report present, skipping... No methylation extractor report present, skipping... Wrote Bismark project summary to >> bismark_summary_report.html << [bam_sort_core] merging from 56 files and 28 in-memory blocks...