Output will be written into the directory: /home/shared/8TB_HDD_01/sam/gitrepos/ceasmallr/output/02.10-bismark-deduplication/ Processing paired-end Bismark output file(s) (SAM format): CF06-CM02-Larvae_R1_001.fastp-trim.20220827_bismark_bt2_pe.bam If there are several alignments to a single position in the genome the first alignment will be chosen. Since the input files are not in any way sorted this is a near-enough random selection of reads. Checking file >>CF06-CM02-Larvae_R1_001.fastp-trim.20220827_bismark_bt2_pe.bam<< for signs of file truncation... samtools view: writing to standard output failed: Broken pipe samtools view: error closing standard output: -1 Now testing Bismark result file CF06-CM02-Larvae_R1_001.fastp-trim.20220827_bismark_bt2_pe.bam for positional sorting (which would be bad...) ...passed! Output file is: CF06-CM02-Larvae_R1_001.fastp-trim.20220827_bismark_bt2_pe.deduplicated.bam Total number of alignments analysed in CF06-CM02-Larvae_R1_001.fastp-trim.20220827_bismark_bt2_pe.bam: 24333642 Total number duplicated alignments removed: 6520652 (26.80%) Duplicated alignments were found at: 4799857 different position(s) Total count of deduplicated leftover sequences: 17812990 (73.20% of total) skipping header line: @HD VN:1.0 SO:unsorted skipping header line: @SQ SN:NC_035780.1 LN:65668440 skipping header line: @SQ SN:NC_035781.1 LN:61752955 skipping header line: @SQ SN:NC_035782.1 LN:77061148 skipping header line: @SQ SN:NC_035783.1 LN:59691872 skipping header line: @SQ SN:NC_035784.1 LN:98698416 skipping header line: @SQ SN:NC_035785.1 LN:51258098 skipping header line: @SQ SN:NC_035786.1 LN:57830854 skipping header line: @SQ SN:NC_035787.1 LN:75944018 skipping header line: @SQ SN:NC_035788.1 LN:104168038 skipping header line: @SQ SN:NC_035789.1 LN:32650045 skipping header line: @SQ SN:NC_007175.2 LN:17244 skipping header line: @PG ID:Bismark VN:v0.24.0 CL:"bismark --path_to_bowtie2 /home/shared/bowtie2-2.4.4-linux-x86_64 --genome /home/shared/8TB_HDD_01/sam/gitrepos/ceasmallr/data/Cvirginica_v300 --score_min L,0,-0.6 --parallel 2 --non_directional --samtools_path /home/shared/samtools-1.12 --gzip -p 4 -1 /home/shared/8TB_HDD_01/sam/gitrepos/ceasmallr/output/02.00-bismark-bowtie2-alignment/trimmed-fastqs/CF06-CM02-Larvae_R1_001.fastp-trim.20220827.fq.gz -2 /home/shared/8TB_HDD_01/sam/gitrepos/ceasmallr/output/02.00-bismark-bowtie2-alignment/trimmed-fastqs/CF06-CM02-Larvae_R2_001.fastp-trim.20220827.fq.gz --output_dir /home/shared/8TB_HDD_01/sam/gitrepos/ceasmallr/output/02.00-bismark-bowtie2-alignment" skipping header line: @PG ID:samtools PN:samtools PP:Bismark VN:1.12 CL:/home/shared/samtools-1.12/samtools view -bSh - skipping header line: @PG ID:samtools.1 PN:samtools PP:samtools VN:1.12 CL:/home/shared/samtools-1.12/samtools view -h /home/shared/8TB_HDD_01/sam/gitrepos/ceasmallr/output/02.00-bismark-bowtie2-alignment/CF06-CM02-Larvae_R1_001.fastp-trim.20220827.fq.gz.temp.1.gz_bismark_bt2_pe.bam skipping header line: @PG ID:samtools.2 PN:samtools PP:samtools.1 VN:1.12 CL:/home/shared/samtools-1.12/samtools view -bSh - skipping header line: @PG ID:samtools.3 PN:samtools PP:samtools.2 VN:1.12 CL:/home/shared/samtools-1.12/samtools view -h --threads 1 CF06-CM02-Larvae_R1_001.fastp-trim.20220827_bismark_bt2_pe.bam samtools view: writing to standard output failed: Broken pipe samtools view: error closing standard output: -1