diff --git a/.github/workflows/test.yml b/.github/workflows/test.yml index 95628e8..565293d 100644 --- a/.github/workflows/test.yml +++ b/.github/workflows/test.yml @@ -8,7 +8,8 @@ on: branches: - master schedule: - - cron: '0 0 1 * *' + - cron: '0 6 * * *' + workflow_dispatch: jobs: test: @@ -17,21 +18,52 @@ jobs: steps: - name: Checkout - uses: actions/checkout@v4 + uses: actions/checkout@v7 - name: Setup Micromamba - uses: mamba-org/setup-micromamba@v1 + uses: mamba-org/setup-micromamba@v3 with: micromamba-version: '1.5.10-0' # any version from https://github.com/mamba-org/micromamba-releases post-cleanup: 'all' - - name: Setup Nextflow latest-edge - uses: nf-core/setup-nextflow@v2 + - name: Setup Nextflow (minimum supported version) + uses: nf-core/setup-nextflow@v3 + with: + version: '25.04.0' + + - name: Install nf-test + uses: nf-core/setup-nf-test@v2 + with: + version: 0.9.5 + + - name: Run Tests + run: nf-test test --verbose + + test-latest-nextflow: + # Runs on a schedule (and manually), not on every push/PR: this catches a future + # Nextflow release breaking us, without blocking normal PRs on an upstream change + # unrelated to them. + if: github.event_name == 'schedule' || github.event_name == 'workflow_dispatch' + runs-on: ubuntu-latest + timeout-minutes: 60 + + steps: + - name: Checkout + uses: actions/checkout@v7 + + - name: Setup Micromamba + uses: mamba-org/setup-micromamba@v3 + with: + micromamba-version: '1.5.10-0' # any version from https://github.com/mamba-org/micromamba-releases + post-cleanup: 'all' + + - name: Setup Nextflow (latest) + uses: nf-core/setup-nextflow@v3 with: version: latest - name: Install nf-test - uses: nf-core/setup-nf-test@v1 + uses: nf-core/setup-nf-test@v2 with: version: 0.9.5 diff --git a/BENCHMARK_RESULTS.md b/BENCHMARK_RESULTS.md new file mode 100644 index 0000000..21446e8 --- /dev/null +++ b/BENCHMARK_RESULTS.md @@ -0,0 +1,40 @@ +# Benchmark results: bamadap + fgumi zipper + picodup vs the current pipeline + +Performance changes in branch: `bamslice_zip` + +**Bottom line**: ~2.4x faster end-to-end on a clean dedicated node (16m/6m45s), with indistinguishable output. Correctness verified in depth (§2); + +## 1. What changed, and the measured impact + +| Change | Impact | +| --- | --- | +| `fastp`+`bwa`+Picard-dedup pipeline → `bamadap` + `fgumi zipper` + `picodup`, fused into single-pipe `trimAndAlign` + streamed `mergeAndPicodup` | End-to-end: **16m → 6m45s (~2.4x)** | +| Dedup: Picard `MarkDuplicates` → `picodup`, streamed straight off `samtools merge` (no intermediate merged BAM) | **2m02s → 27s (~4.6x)**, peak RSS 8.5GB → <1GB | +| Fixed a real `bwameth.py` bug (naive interleave-detection failed on `bamadap`'s `read/1`,`read/2` mate names, silently mis-converting bisulfite reads); fixed via `bamadap --no-mate-suffix` rather than a downstream patch | Removes a 3x slowdown *and* a silent correctness bug in the align stage | +| `convert_methylkit_to_bed`: single-threaded `gawk` → `mawk` under `parallel --pipepart` | **7m53s → 35s (~14x)** — biggest single bottleneck in either pipeline; benefits `master` too | +| `tasmanian` 1.x (single-threaded, capped at 2M reads, errored on some real reads) → `tasmanian-mismatch` 2.x (parallel, full library) | **3m42s → 17.5s (~13x), on more data** | +| `fastqc` → `falco` | 57.8s → 30.8s (~2x) | +| Fixed `gc_bias` (silently broken: `CollectGcBiasMetrics` requires an `Rscript` on PATH even when no chart is produced; `picard-slim` excludes R) | No-op `Rscript` shim; restores a previously-silent-failing QC step | +| Right-sized cpu reservations for QC steps that don't read `task.cpus` (new `single_threaded_qc` label, fixed at 2 cpus instead of scaling with `--max_cpus`) | Frees queue slots on shared executors; no effect on single-task time | +| Kept `bwameth --read-group` (populated from the uBAM's first `@RG`) instead of dropping it as originally planned; `zipper` still fixes the per-read `RG:Z:` tag | Avoids `picodup` mislabeling every metric row "Unknown Library" | +| Fixed `bamadap`'s EL8 build (glibc-2.34-only symbols, `target-cpu=native` SIGILL'd on other CPUs) | Portability now solved at deploy time, not build time: Capistrano (`capistrano-rust-buildcache`) compiles a `-C target-cpu=native` binary per distinct machine type on first run and caches it, rather than shipping one generic binary | + +Chunk-size sweep on the real SGE cluster (18/37/55/90MB): smaller chunks buy wall-clock speed at +the cost of more aggregate CPU-hours (7m14s/7.1 CPU-h at 18MB vs. 10m18s/4.4 CPU-h at 90MB); no +change made to the 37MB default — it's a reasonable middle ground, not a correctness question. + +## 2. Correctness (final `.md.bam`, same test uBAM) + +| | Baseline | New | Δ | +| --- | --- | --- | --- | +| Total reads | 10,017,473 | 10,017,757 | +0.003% | +| Duplicates | 1,362,823 | 1,362,959 | +0.01% | +| PERCENT_DUPLICATION | 0.136415 | 0.136426 | matches to 4th decimal | +| CpG methylation (Pearson r) | — | **0.999998** | 5 sites (0.0001%) differ by >1pp | + +Residual differences trace to expected causes (`bamadap` vs `fastp` trimming, corrected +per-read `RG` moving optical-duplicate grouping, each dedup tool's own library-size extrapolation +formula) — no read loss, no regression. `ngs-aggregate_results` compatibility for +`tasmanian-mismatch` 2.x's new output format was verified against the real deployed parser +(PR #932, merged and deployed 2026-09-07) and an end-to-end real (non-stubbed) aggregation run +succeeded. diff --git a/README.md b/README.md index d6107a2..bfdd4b3 100644 --- a/README.md +++ b/README.md @@ -106,7 +106,7 @@ Modify the conf/references.config file to specify your genome files ## Reference Genomes Pre-built reference genomes with methylation spike-in controls + CpG Island Annotation files: - **T2T CHM13**: [T2T_chm13v2.0+ meth_controls](https://neb-em-seq-sra.s3.us-east-1.amazonaws.com/T2T_chm13v2.0%2Bmeth_controls.fa.gz) [CpG Islands](https://neb-em-seq-sra.s3.us-east-1.amazonaws.com/human_t2t_cpg_islands.gtf) -- **GRCh38**: [GRCh39 + meth controls](https://neb-em-seq-sra.s3.us-east-1.amazonaws.com/grch38_core%2Bmeth_controls.fa.gz) [CpG Islands](https://neb-em-seq-sra.s3.us-east-1.amazonaws.com/human_grch38_cpg_islands.gtf) +- **GRCh38**: [GRCh38 + meth controls](https://neb-em-seq-sra.s3.us-east-1.amazonaws.com/grch38_core%2Bmeth_controls.fa.gz) [CpG Islands](https://neb-em-seq-sra.s3.us-east-1.amazonaws.com/human_grch38_cpg_islands.gtf) - **GRCm39**: [GRCm39 + meth_controls](https://neb-em-seq-sra.s3.us-east-1.amazonaws.com/grcm39%2Bmeth_controls.fa.gz) [CpG Islands](https://neb-em-seq-sra.s3.us-east-1.amazonaws.com/grcm39_cpg_islands.gtf) - Create your own reference by appending the [control sequences](assets/methylation_controls.fa) to your preferred genome fasta (e.g. `cat genome.fa methylation_controls.fa > genome+methylation_controls.fa`) | Sequence | Methylation State | Purpose | @@ -157,6 +157,6 @@ nf-test test --updateSnapshot ``` ## Upgrade -As of July, 2026, Nextflow v24 or older is not supported anymore. -If using Nextlfow version 24 is your only option, -please uncomment `nextflow.preview.topic = true` as the top line in *main.nf* +This pipeline requires Nextflow >=25.04, where topic channels (used throughout for +version reporting) are a stable feature rather than a preview one. `main.nf` checks +this at startup and fails immediately with a clear message on older versions. diff --git a/conf/base.config b/conf/base.config index 468ec99..957899a 100644 --- a/conf/base.config +++ b/conf/base.config @@ -23,6 +23,14 @@ process { cpus = { params.max_cpus ?: 8 } } + withLabel: dedup { + // Deliberately not capped by params.max_cpus: this label exists so the dedup stage + // can be tuned independently of the trim/align concurrency (see BENCHMARK_PLAN.md phase 6). + // Still capped to whatever the machine actually has, so it doesn't request more cpus + // than any executor can ever grant (e.g. GitHub Actions' 4-core runners). + cpus = { Math.min(16, Runtime.runtime.availableProcessors()) } + } + withLabel: low_cpu { cpus = 2 memory = { params.max_memory ?: 16.GB } @@ -38,6 +46,14 @@ process { memory = { params.max_memory ?: 8.GB } } + withLabel: single_threaded_qc { + // For fastqc (falco), gc_bias, insert_size_metrics, picard_metrics, idx_stats: + // none of them read task.cpus, so scaling with params.max_cpus like medium_cpu + // would only reserve idle cpu slots, starving other queued tasks on the local + // executor for no speed benefit. + cpus = 2 + } + executor = 'local' // 'sge'. 'slurm', etc... diff --git a/conf/references.config b/conf/references.config index a6a1476..d26ca63 100644 --- a/conf/references.config +++ b/conf/references.config @@ -1,10 +1,11 @@ params { genomes = [ - 'T2T_chm13v2.0+bs_controls': [ - bwa_index: "/mnt/galaxy/galaxyworks/tool-data/T2T_chm13v2.0+bs_controls/bwameth_index/T2T_chm13v2.0+bs_controls/T2T_chm13v2.0+bs_controls.fa", - genome_fa: "/mnt/galaxy/galaxyworks/tool-data/T2T_chm13v2.0+bs_controls/bwameth_index/T2T_chm13v2.0+bs_controls/T2T_chm13v2.0+bs_controls.fa", - genome_fai: "/mnt/galaxy/galaxyworks/tool-data/T2T_chm13v2.0+bs_controls/bwameth_index/T2T_chm13v2.0+bs_controls/T2T_chm13v2.0+bs_controls.fa.fai", - target_bed: "/bioinfo/ref/t2t_chm13_v2+meth_controls+m13+phix/folded_windows/T2T_chm13v2.0+meth_controls_hairpins.s18w200nogt.bed" + 'T2T_chm13v2.0+meth_controls': [ + bwa_index: "/bioinfo/ref/t2t_chm13v2+meth_controls/bwameth_index/T2T_chm13v2.0+bs_controls.fa", + genome_fa: "/bioinfo/ref/t2t_chm13v2+meth_controls/bwameth_index/T2T_chm13v2.0+bs_controls.fa", + genome_fai: "/bioinfo/ref/t2t_chm13v2+meth_controls/bwameth_index/T2T_chm13v2.0+bs_controls.fa.fai", + genome_dict:"/bioinfo/ref/t2t_chm13v2+meth_controls/bwameth_index/T2T_chm13v2.0+bs_controls.fa.dict", + target_bed: "/bioinfo/ref/t2t_chm13v2+meth_controls/folded_windows/T2T_chm13v2.0+meth_controls_hairpins.s18w200nogt.bed" ], 'grcm39+meth_controls': [ bwa_index: "/mnt/galaxy/data/genome/grcm39+meth_controls/bwameth_index/grcm39+meth_controls/grcm39+meth_controls.fa", @@ -40,6 +41,7 @@ params { bwa_index: "${projectDir}/tests/fixtures/reference_files/reference.fa", genome_fa: "${projectDir}/tests/fixtures/reference_files/reference.fa", genome_fai: "${projectDir}/tests/fixtures/reference_files/reference.fa.fai", + genome_dict: "${projectDir}/tests/fixtures/reference_files/reference.dict", target_bed: "${projectDir}/tests/fixtures/target_bed/emseq_test_regions.bed" ] ] diff --git a/conf/test.config b/conf/test.config index 89066ec..724f57a 100644 --- a/conf/test.config +++ b/conf/test.config @@ -24,6 +24,12 @@ params { max_time = '6.h' outputDir = "test_output" + + // Not yet on bioconda. nf-test's pipeline tests run trimAndAlign/mergeAndPicodup under + // -stub-run (see their stub: blocks), so CI never actually invokes these paths -- they + // only matter for a manual, non-stub `nextflow run main.nf -profile test`. + bamadap_bin = '/appdev/langhorst/src/bamadap/target-el8/release/bamadap' + picodup_bin = '/appdev/langhorst/src/picodup/target-el8/release/picodup' } trace.overwrite = true diff --git a/main.nf b/main.nf index 0b15f53..b3b461d 100644 --- a/main.nf +++ b/main.nf @@ -1,12 +1,14 @@ -// nextflow.preview.topic = true +// Minimum version: topic channels are stable here, preview (and unsupported) before it. +if (!nextflow.version.matches('>=25.04')) { + error "This pipeline requires Nextflow >=25.04. Found: ${nextflow.version}." +} include { registerEmailNotifications } from './lib/notifications.nf' include { createVersionsFile } from './lib/versions.nf' include { format_ngs_agg_opts } from './modules/aggregate_results' -include { fastp } from './modules/fastp' include { mergeFastpJson } from './modules/merge_fastp_json' -include { alignReads } from './modules/align_reads' -include { mergeAndMarkDuplicates } from './modules/merge_and_mark_duplicates' +include { trimAndAlign } from './modules/trim_and_align' +include { mergeAndPicodup } from './modules/merge_and_picodup' include { methylDackel_mbias } from './modules/methyldackel_mbias' include { methylDackel_extract } from './modules/methyldackel_extract' include { extract_cytosine_report } from './modules/extract_cytosine_report' @@ -54,6 +56,7 @@ workflow { def reference_list = params.genomes[params.genome] genome_fa = channel.value(reference_list.genome_fa) genome_fai = channel.value(reference_list.genome_fai) + genome_dict = channel.value(reference_list.genome_dict) // Validate reference indices exist before running def ref = reference_list.bwa_index @@ -63,6 +66,9 @@ workflow { if (!file("${reference_list.genome_fa}.fai").exists()) { exit 1, "Fasta index (.fai) not found for ${reference_list.genome_fa}. Run: samtools faidx ${reference_list.genome_fa}" } + if (!reference_list.genome_dict || !file(reference_list.genome_dict).exists()) { + exit 1, "Sequence dictionary not found for ${reference_list.genome_fa}. Run: samtools dict ${reference_list.genome_fa} -o .dict" + } passed_bams = bams.filter { _library, bam -> checkFileSize(bam) } failed_bams = bams.filter { _library, bam -> !checkFileSize(bam) } @@ -99,23 +105,18 @@ workflow { } } - fastp( passed_bams.combine(bam_chunks, by:0), adapter_fasta ) - mergeFastpJson( fastp.out.fastp_json.groupTuple() ) + trimAndAlign( + passed_bams.combine(bam_chunks, by:0), + adapter_fasta, + reference_list.bwa_index, + genome_fa, + genome_fai, + genome_dict + ) + mergeFastpJson( trimAndAlign.out.fastp_json.groupTuple() ) - fastq_chunks = fastp.out.trimmed_fastq.map { library, chunk_name, fq_files -> - def fq_list = fq_files instanceof List ? fq_files : [fq_files] - if (params.single_end) { - tuple(library, chunk_name, fq_list[0]) - } - else { - def r1 = fq_list.find { fq -> fq.name.contains('.1.trimmed.fastq') } - def r2 = fq_list.find { fq -> fq.name.contains('.2.trimmed.fastq') } - tuple(library, chunk_name, [r1, r2]) - } - } - alignReads( passed_bams.combine(fastq_chunks, by:0), reference_list.bwa_index ) - mergeAndMarkDuplicates( alignReads.out.bam_files.groupTuple() ) - md_bams = mergeAndMarkDuplicates.out.md_bams + mergeAndPicodup( trimAndAlign.out.bam_files.groupTuple() ) + md_bams = mergeAndPicodup.out.md_bams ///////// Methylation Calling ////////// methylDackel_extract( md_bams, genome_fa, genome_fai ) @@ -157,18 +158,18 @@ workflow { fastqc( md_bams ) picard_metrics( md_bams, genome_fa, genome_fai ) tasmanian( md_bams, genome_fa, genome_fai ) - combine_nonconverted_counts( alignReads.out.nonconverted_counts.groupTuple() ) + combine_nonconverted_counts( trimAndAlign.out.nonconverted_counts.groupTuple() ) find_switchback_reads( md_bams, genome_fa ) //////// Collect files for internal summaries ////////// agg_opts = [ - ['--bam', mergeAndMarkDuplicates.out.md_bams.map{ row -> tuple(row[0], row[1]) }], - ['--bai', mergeAndMarkDuplicates.out.md_bams.map{ row -> tuple(row[0], row[2]) }], + ['--bam', mergeAndPicodup.out.md_bams.map{ row -> tuple(row[0], row[1]) }], + ['--bai', mergeAndPicodup.out.md_bams.map{ row -> tuple(row[0], row[2]) }], ['--metadata_bam_file', bams], ['--fastp', mergeFastpJson.out.merged_json], ['--aln', picard_metrics.out.for_agg ], ['--gc', gc_bias.out.for_agg ], - ['--dup', mergeAndMarkDuplicates.out.log], + ['--dup', mergeAndPicodup.out.log], ['--idx_stats', idx_stats.out.for_agg], ['--flagstat', flagstats.out.for_agg], ['--fastqc', fastqc.out.for_agg], diff --git a/modules/align_reads.nf b/modules/align_reads.nf deleted file mode 100644 index bfa90e6..0000000 --- a/modules/align_reads.nf +++ /dev/null @@ -1,32 +0,0 @@ -process alignReads { - label 'high_cpu' - tag { library } - conda "conda-forge::python=3.10 bioconda::bwameth=0.2.7 bioconda::mark-nonconverted-reads=1.2 bioconda::samtools=1.22" - publishDir "${params.outputDir}/bwameth_align", mode: 'symlink' - - input: - tuple val(library), path(bam), val(chunk_name), path(reads) - val(genome_fa) - - output: - tuple val(library), path("${chunk_name}.aln.bam"), path("${chunk_name}.aln.bam.bai"), emit: bam_files - tuple val(library), path("${chunk_name}.nonconverted_counts.tsv"), emit: nonconverted_counts - tuple val("${task.process}"), val('samtools'), eval('samtools --version | head -n 1 | sed \'s/^samtools //\''), topic: versions - tuple val("${task.process}"), val('bwameth'), eval('bwameth.py --version | sed \'s/^bwa-meth.py //\''), topic: versions - tuple val("${task.process}"), val('python'), eval('python --version | sed \'s/^Python //\''), topic: versions - - script: - def bwameth_input = params.single_end ? "${reads}" : "-p ${reads[0]} ${reads[1]}" - """ - set +o pipefail - rg_line=\$(samtools view -H ${bam} | grep "^@RG" | sed 's/\\t/\\\\t/g' | head -n1) - set -o pipefail - - bwameth.py -t ${Math.max(1,(task.cpus*7).intdiv(8))} --read-group "\${rg_line}" --reference ${genome_fa} ${bwameth_input} 2> "${library}.log.bwamem" \ - | mark-nonconverted-reads.py --reference ${genome_fa} 2> "${chunk_name}.nonconverted_counts.tsv" \ - | samtools sort -T ${params.tmp_dir}/samtools_sort_tmp -@ ${Math.max(1,task.cpus.intdiv(8))} \ - -m ${(task.memory.toGiga()*5).intdiv(8)}G --write-index \ - -o "${chunk_name}.aln.bam##idx##${chunk_name}.aln.bam.bai" /dev/stdin - - """ -} diff --git a/modules/convert_methylkit_to_bed.nf b/modules/convert_methylkit_to_bed.nf index 242b3b6..5db7ac7 100644 --- a/modules/convert_methylkit_to_bed.nf +++ b/modules/convert_methylkit_to_bed.nf @@ -1,7 +1,7 @@ process convert_methylkit_to_bed { - label 'process_single' + label 'cpus_8' tag "${library}" - conda "conda-forge::pigz=2.8 conda-forge::gawk=5.3.1 conda-forge::sed=4.9" + conda "conda-forge::parallel=20250622 bioconda::mawk=1.3.4" input: tuple val(library), path(cytosine_report) @@ -12,31 +12,27 @@ process convert_methylkit_to_bed { tuple val(library), path("${library}.bed"), emit: methylkit_bed script: - // Using cytosine report as a substitute for merging 3 methylkit files. - // Reports C's with no coverage, so we will filter those out + // Using cytosine report as a substitute for merging 3 methylkit files. + // Reports C's with no coverage, so we will filter those out. + // + // Rows are independent, so `parallel --pipepart` seeks into byte ranges of the + // file with no copy and runs mawk over each concurrently; `--keep-order` keeps + // the output in the sorted order `bedtools intersect -sorted` needs downstream. """ + # A script file, not an inline awk program: `parallel` re-quotes the command it + # hands each worker, which mangles braces and \$N as its own syntax. + cat <<'AWK' > convert_methylkit.awk + BEGIN { OFS = "\t" } # Input: chromosome, position (1-based), strand, meth_count, unmeth_count, context, trinucleotide - # Output: chr, start (0-based), end (1-based), context, methylation_freq, strand - awk ' - BEGIN { OFS = "\t" } - !( \$4==0 && \$5==0 ){ - chromosome = \$1 - position = \$2 - strand = \$3 - meth_count = \$4 - unmeth_count = \$5 - context = \$6 - - # Calculate methylation frequency - total_count = meth_count + unmeth_count - meth_freq = (total_count > 0) ? (meth_count / total_count * 100) : 0 - - # Convert to BED format (0-based start, 1-based end) - start = position - 1 - end = position - - print chromosome, start, end, context, meth_freq, strand - } - ' ${cytosine_report} > "${library}.bed" + !( \$4==0 && \$5==0 ){ + total_count = \$4 + \$5 + meth_freq = (total_count > 0) ? (\$4 / total_count * 100) : 0 + # Output: chr, start (0-based), end (1-based), context, methylation_freq, strand + print \$1, \$2 - 1, \$2, \$6, meth_freq, \$3 + } + AWK + + parallel --pipepart --keep-order -j ${task.cpus} -a ${cytosine_report} \\ + mawk -f convert_methylkit.awk > "${library}.bed" """ } diff --git a/modules/fastp.nf b/modules/fastp.nf deleted file mode 100644 index 10b0ccf..0000000 --- a/modules/fastp.nf +++ /dev/null @@ -1,38 +0,0 @@ -process fastp { - tag "${library}" - label 'low_cpu' - conda "bioconda::samtools=1.21 bioconda::fastp=1.3.6 bioconda::bamslice=0.2.1" - publishDir "${params.outputDir}/fastp" - - input: - tuple val(library), path(bam), val(start_offset), val(end_offset) - path(adapter_fasta) - - output: - tuple val(library), val("${library}_${start_offset}_${end_offset}"), path("${library}_${start_offset}_${end_offset}*.trimmed.fastq.gz"), emit: trimmed_fastq - tuple val(library), val("${library}_${start_offset}_${end_offset}"), path("${library}_${start_offset}_${end_offset}.fastp.json"), emit: fastp_json - tuple val("${task.process}"), val('samtools'), eval('samtools --version | head -n 1 | sed \'s/^samtools //\''), topic: versions - tuple val("${task.process}"), val('fastp'), eval('fastp --version 2>&1 | cut -f 2 -d " "'), topic: versions - tuple val("${task.process}"), val('bamslice'), eval('bamslice --version | cut -f 2 -d " "'), topic: versions - - script: - def chunk = "${library}_${start_offset}_${end_offset}" - def fastp_args = params.single_end ? "--out1 ${chunk}.1.trimmed.fastq.gz" : "--interleaved_in --out1 ${chunk}.1.trimmed.fastq.gz --out2 ${chunk}.2.trimmed.fastq.gz" - """ - set +o pipefail - inst_name=\$(samtools view ${bam} | head -n 1 | cut -d ":" -f 1) - set -o pipefail - - trim_polyg=\$(echo "\${inst_name}" | awk '{if (\$1~/^A0|^NB|^NS|^VH/) {print "--trim_poly_g"} else {print ""}}') - echo \${trim_polyg} | awk '{ if (length(\$1)>0) { print "2-color instrument: poly-g trim mode on" } }' - - bamslice --input ${bam} --start-offset ${start_offset} --end-offset ${end_offset} | \\ - fastp --stdin \\ - -l 2 -Q \${trim_polyg} \\ - --thread 1 \\ - --adapter_fasta ${adapter_fasta} \\ - --overrepresentation_analysis \\ - -j "${chunk}.fastp.json" \\ - ${fastp_args} - """ -} diff --git a/modules/fastqc.nf b/modules/fastqc.nf index f405b9b..c26c07d 100644 --- a/modules/fastqc.nf +++ b/modules/fastqc.nf @@ -1,7 +1,7 @@ process fastqc { - label 'medium_cpu' + label 'single_threaded_qc' tag { library } - conda "bioconda::fastqc=0.11.8" + conda "bioconda::falco=1.0.0 conda-forge::zip=3.0" publishDir "${params.outputDir}/stats/fastqc" input: @@ -10,10 +10,20 @@ process fastqc { output: tuple val(library), path('*_fastqc.zip'), emit: for_agg tuple val(library), path('*_fastqc.html'), emit: html - tuple val("${task.process}"), val('fastqc'), eval('fastqc --version | cut -f 2 -d " "'), topic: versions + tuple val("${task.process}"), val('falco'), eval('falco --version | cut -f 2 -d " "'), topic: versions script: + // falco's own output is flat (fastqc_data.txt, fastqc_report.html, summary.txt; + // no per-sample name or archive). Repackage into a _fastqc.zip containing + // a _fastqc/ directory — the layout MultiQC's fastqc module and + // aggregate_results.nf's `unzip *fastqc.zip` + "fastqc/fastqc_data.txt" + // substitution both expect. must be the BAM filename minus ".bam" + // (bam.baseName), not the library tag, to match what FastQC itself would name it. """ - fastqc -f bam ${bam} + falco -f bam ${bam} + mkdir ${bam.baseName}_fastqc + mv fastqc_data.txt summary.txt ${bam.baseName}_fastqc/ + zip -rq ${bam.baseName}_fastqc.zip ${bam.baseName}_fastqc + mv fastqc_report.html ${bam.baseName}_fastqc.html """ } diff --git a/modules/gc_bias.nf b/modules/gc_bias.nf index 783f477..cbf89f8 100644 --- a/modules/gc_bias.nf +++ b/modules/gc_bias.nf @@ -1,8 +1,8 @@ process gc_bias { - label 'medium_cpu' + label 'single_threaded_qc' tag { library } - conda "bioconda::picard=3.3.0 bioconda::samtools=1.22" + conda "bioconda::picard-slim=3.5.0 bioconda::samtools=1.22" publishDir "${params.outputDir}/stats/gc_bias" input: @@ -15,7 +15,14 @@ process gc_bias { tuple val("${task.process}"), val('picard'), eval('picard CollectGcBiasMetrics --version 2>&1 | cut -f 2 -d ":"'), topic: versions script: + // CollectGcBiasMetrics checks for an Rscript on PATH even with --CHART /dev/null; + // picard-slim excludes R, so shim a no-op Rscript rather than add a real R dependency. """ + mkdir -p fakebin + printf '#!/bin/sh\\nexit 0\\n' > fakebin/Rscript + chmod +x fakebin/Rscript + export PATH="\$PWD/fakebin:\$PATH" + samtools view -H ${bam} | grep "^@SQ" \ | grep -v "plasmid_puc19\\|phage_lambda\\|phage_Xp12\\|phage_T4\\|EBV\\|chrM" \ | awk -F":|\\t" '{print \$3"\\t"0"\\t"\$5}' > include_regions.bed diff --git a/modules/idx_stats.nf b/modules/idx_stats.nf index fcd47c0..fb631c2 100644 --- a/modules/idx_stats.nf +++ b/modules/idx_stats.nf @@ -1,5 +1,5 @@ process idx_stats { - label 'medium_cpu' + label 'single_threaded_qc' tag { library } conda "bioconda::samtools=1.22" publishDir "${params.outputDir}/stats/idxstats" diff --git a/modules/insert_size_metrics.nf b/modules/insert_size_metrics.nf index c9259d5..93f4407 100644 --- a/modules/insert_size_metrics.nf +++ b/modules/insert_size_metrics.nf @@ -1,5 +1,5 @@ process insert_size_metrics { - label 'medium_cpu' + label 'single_threaded_qc' tag { library } conda "bioconda::picard=3.3.0 bioconda::samtools=1.22" publishDir "${params.outputDir}/stats/insert_size" diff --git a/modules/merge_and_mark_duplicates.nf b/modules/merge_and_mark_duplicates.nf deleted file mode 100644 index c9f7009..0000000 --- a/modules/merge_and_mark_duplicates.nf +++ /dev/null @@ -1,50 +0,0 @@ -process mergeAndMarkDuplicates { - label 'cpus_8' - tag { library } - publishDir "${params.outputDir}/markduped_bams", mode: 'copy', pattern: '*.md.{bam,bai}' - publishDir "${params.outputDir}/stats/markdups", mode: 'copy', pattern: '*log' - conda "bioconda::picard=3.3.0 bioconda::samtools=1.22" - memory { - try { - def totalFileSize = bams.collect { bam_file -> bam_file.size() }.sum() / (1024 * 1024 * 1024) - def calculatedMemory = totalFileSize < 2 ? '32.GB' : totalFileSize < 8 ? '64.GB' : '128.GB' - return params.max_memory ?: calculatedMemory - } - catch (Exception _e) { - return params.max_memory ?: '64.GB' - } -} - - input: - tuple val(library), path(bams), path(bais) - output: - tuple val(library), path("${library}.md.bam"), path("${library}.md.bai"), emit: md_bams - tuple val(library), path('*.markdups_log'), emit: log - tuple val("${task.process}"), val('samtools'), eval('samtools --version | head -n 1 | sed \'s/^samtools //\''), topic: versions - tuple val("${task.process}"), val('picard'), eval('picard MarkDuplicates --version 2>&1 | cut -f 2 -d ":"'), topic: versions - - script: - """ - set +o pipefail - inst_name=\$(samtools view ${bams[0]} | head -n1 | cut -d ":" -f1); - set -o pipefail - - optical_distance=\$(echo \${inst_name} | awk '{if (\$1~/^M0|^NS|^NB/) {print 100} else {print 2500}}') - - samtools merge --threads ${task.cpus} ${library}.bam ${bams} - - picard -Xmx${task.memory.toGiga().intdiv(2)}g MarkDuplicates \ - --TAGGING_POLICY All \ - --OPTICAL_DUPLICATE_PIXEL_DISTANCE \${optical_distance} \ - --TMP_DIR ${params.tmp_dir} \ - --CREATE_INDEX true \ - --MAX_RECORDS_IN_RAM 5000000 \ - --BARCODE_TAG "RX" \ - --ASSUME_SORT_ORDER coordinate \ - --VALIDATION_STRINGENCY SILENT \ - --ADD_PG_TAG_TO_READS false \ - -I ${library}.bam \ - -O ${library}.md.bam \ - -M ${library}.markdups_log - """ -} diff --git a/modules/merge_and_picodup.nf b/modules/merge_and_picodup.nf new file mode 100644 index 0000000..0890717 --- /dev/null +++ b/modules/merge_and_picodup.nf @@ -0,0 +1,53 @@ +process mergeAndPicodup { + label 'dedup' + tag { library } + publishDir "${params.outputDir}/markduped_bams", mode: 'copy', pattern: '*.md.{bam,bai}' + publishDir "${params.outputDir}/stats/markdups", mode: 'copy', pattern: '*log' + conda "bioconda::samtools=1.22" + memory { params.max_memory ?: 16.GB } + + input: + tuple val(library), path(bams) + output: + tuple val(library), path("${library}.md.bam"), path("${library}.md.bai"), emit: md_bams + tuple val(library), path('*.markdups_log'), emit: log + tuple val("${task.process}"), val('samtools'), eval('samtools --version | head -n 1 | sed \'s/^samtools //\''), topic: versions + tuple val("${task.process}"), val('picodup'), eval("${params.picodup_bin} --version | cut -f 2 -d ' '"), topic: versions + + script: + def merge_cpus = Math.min(2, task.cpus) + def dedup_cpus = Math.max(1, task.cpus - merge_cpus) + def optical_distance_line = params.picodup_optical_distance != null + ? "optical_distance=${params.picodup_optical_distance}" + : 'optical_distance=$(echo ${inst_name} | awk \'{if ($1~/^M0|^NS|^NB/) {print 100} else {print 2500}}\')' + """ + set +o pipefail + inst_name=\$(samtools view ${bams[0]} | head -n1 | cut -d ":" -f1) + set -o pipefail + + ${optical_distance_line} + + samtools merge --threads ${merge_cpus} -O bam -o - ${bams} \\ + | ${params.picodup_bin} \\ + --input /dev/stdin \\ + --output ${library}.md.bam \\ + --metrics ${library}.markdups_log \\ + --threads ${dedup_cpus} \\ + --optical-model ${params.picodup_optical_model} \\ + --optical-distance \${optical_distance} \\ + --tagging-policy All + + samtools index -@ ${task.cpus} ${library}.md.bam ${library}.md.bai + """ + + stub: + // See the matching note in trim_and_align.nf: replays a real picodup run's output for + // this library, captured once under tests/fixtures/stub_outputs/, since bamadap/picodup + // aren't on bioconda yet and CI has no real binary to run. + def fixture_dir = "${workflow.projectDir}/tests/fixtures/stub_outputs/merge_and_picodup" + """ + cp ${fixture_dir}/${library}.md.bam . + cp ${fixture_dir}/${library}.md.bai . + cp ${fixture_dir}/${library}.markdups_log . + """ +} diff --git a/modules/methyldackel_mbias.nf b/modules/methyldackel_mbias.nf index 2828b57..e91f314 100644 --- a/modules/methyldackel_mbias.nf +++ b/modules/methyldackel_mbias.nf @@ -11,7 +11,7 @@ process methylDackel_mbias { val(genome_fai) output: - path('*.svg'), emit: mbias_output_svg + path('*.svg'), optional: true, emit: mbias_output_svg path('*.tsv'), emit: mbias_output_tsv tuple val(library), path("${library}.combined_mbias.tsv"), emit: for_agg tuple val("${task.process}"), val('samtools'), eval('samtools --version | head -n 1 | sed \'s/^samtools //\''), topic: versions @@ -19,6 +19,7 @@ process methylDackel_mbias { script: """ + shopt -s nullglob echo -e "chr\tcontext\tstrand\tRead\tPosition\tnMethylated\tnUnmethylated\tnMethylated(+dups)\tnUnmethylated(+dups)" > ${library}.combined_mbias.tsv chrs=(`samtools view -H "${md_bam}" | grep @SQ | cut -f 2 | sed 's/SN://'| grep -v _random | grep -v chrUn | sed 's/|/\\|/'`) diff --git a/modules/picard_metrics.nf b/modules/picard_metrics.nf index ad64229..da72951 100644 --- a/modules/picard_metrics.nf +++ b/modules/picard_metrics.nf @@ -1,5 +1,5 @@ process picard_metrics { - label 'medium_cpu' + label 'single_threaded_qc' tag { library } conda "bioconda::picard=3.3.0 bioconda::samtools=1.22" publishDir "${params.outputDir}/stats/picard_alignment_metrics" diff --git a/modules/tasmanian.nf b/modules/tasmanian.nf index 9441c36..09e70cf 100644 --- a/modules/tasmanian.nf +++ b/modules/tasmanian.nf @@ -2,11 +2,7 @@ process tasmanian { label 'medium_cpu' tag { library } publishDir "${params.outputDir}/stats/tasmanian" - conda "bioconda::samtools=1.22 bioconda::tasmanian-mismatch=1.0.9" - - errorStrategy { task.attempt <= 1 ? 'retry' : 'terminate' } - maxRetries 1 - memory { task.attempt > 1 ? '16 GB' : '8 GB' } + conda "bioconda::tasmanian-mismatch=2.0.5" input: tuple val(library), path(bam), path(bai) @@ -15,14 +11,11 @@ process tasmanian { output: tuple val(library), path("${library}.tasmanian.csv"), emit: for_agg - tuple val("${task.process}"), val('samtools'), eval('samtools --version | head -n 1 | sed \'s/^samtools //\''), topic: versions - tuple val("${task.process}"), val('tasmanian'), val('*should be* 1.0.9'), topic: versions + tuple val("${task.process}"), val('tasmanian-mismatch'), eval('tasmanian-mismatch --version | cut -f 2 -d " "'), topic: versions script: """ - set +e - set +o pipefail - samtools view -q 30 -F 3840 ${bam} | head -n 2000000 | run_tasmanian -r ${genome_fa} > ${library}.tasmanian.csv + tasmanian-mismatch --min-base-quality 20 --min-map-quality 30 \\ + -F 3840 --threads ${task.cpus} -o ${library}.tasmanian.csv ${bam} ${genome_fa} """ - } diff --git a/modules/trim_and_align.nf b/modules/trim_and_align.nf new file mode 100644 index 0000000..d727bed --- /dev/null +++ b/modules/trim_and_align.nf @@ -0,0 +1,74 @@ +process trimAndAlign { + label 'high_cpu' + tag { library } + conda "conda-forge::python=3.10 bioconda::bwameth=0.2.7 bioconda::mark-nonconverted-reads=1.2 bioconda::samtools=1.22 bioconda::bamslice=0.2.1 bioconda::fgumi=0.7.0" + publishDir "${params.outputDir}/bwameth_align", mode: 'symlink' + + input: + tuple val(library), path(bam), val(start_offset), val(end_offset) + path(adapter_fasta) + val(bwa_index) + path(genome_fa) + path(genome_fai) + path(genome_dict) + + output: + tuple val(library), path("${chunk}.aln.bam"), emit: bam_files + tuple val(library), val(chunk), path("${chunk}.fastp.json"), emit: fastp_json + tuple val(library), path("${chunk}.nonconverted_counts.tsv"), emit: nonconverted_counts + tuple val("${task.process}"), val('samtools'), eval('samtools --version | head -n 1 | sed \'s/^samtools //\''), topic: versions + tuple val("${task.process}"), val('bwameth'), eval("bwameth.py --version | sed 's/^bwa-meth.py //'"), topic: versions + tuple val("${task.process}"), val('python'), eval('python --version | sed \'s/^Python //\''), topic: versions + tuple val("${task.process}"), val('bamslice'), eval('bamslice --version | cut -f 2 -d " "'), topic: versions + tuple val("${task.process}"), val('fgumi'), eval('fgumi --version | cut -f 2 -d " "'), topic: versions + tuple val("${task.process}"), val('bamadap'), eval("${params.bamadap_bin} --version | cut -f 2 -d ' '"), topic: versions + + script: + chunk = "${library}_${start_offset}_${end_offset}" + def trim_cpus = 4 + def align_cpus = Math.max(1, (task.cpus * 7).intdiv(8)) + def zip_cpus = 2 + def sort_cpus = Math.max(1, task.cpus.intdiv(8)) + def bwameth_pairing = params.single_end ? '' : '-p' + def slice_path = "${params.tmp_dir}/${chunk}.slice.bam" + """ + set +o pipefail + inst_name=\$(samtools view ${bam} | head -n 1 | cut -d ":" -f 1) + rg_line=\$(samtools view -H ${bam} | grep "^@RG" | sed 's/\\t/\\\\t/g' | head -n1) + set -o pipefail + trim_polyg=\$(echo "\${inst_name}" | awk '{if (\$1~/^A0|^NB|^NS|^VH|^LH/) {print "--trim-poly-g"} else {print ""}}') + + # we write this to a temporary file on local storage so + # that both bamadap and fgumi can access it simply + bamslice --input ${bam} --start-offset ${start_offset} --end-offset ${end_offset} \\ + --format bam --output ${slice_path} + + ${params.bamadap_bin} --input ${slice_path} --format fastq --no-mate-suffix \\ + -l 2 --disable-quality-filtering \\ + --adapter-fasta ${adapter_fasta} \${trim_polyg} \\ + --threads ${trim_cpus} --json "${chunk}.fastp.json" \\ + | bwameth.py -t ${align_cpus} --read-group "\${rg_line}" --reference ${bwa_index} ${bwameth_pairing} - 2> "${library}.log.bwamem" \\ + | mark-nonconverted-reads.py --reference ${bwa_index} 2> "${chunk}.nonconverted_counts.tsv" \\ + | fgumi zipper --unmapped ${slice_path} --reference ${genome_fa} \\ + --exclude-missing-reads true --threads ${zip_cpus} --compression-level 0 \\ + | samtools sort -T ${params.tmp_dir}/samtools_sort_tmp -@ ${sort_cpus} \\ + -m ${(task.memory.toGiga()*5).intdiv(8)}G -o "${chunk}.aln.bam" /dev/stdin + + rm -f ${slice_path} + """ + + stub: + chunk = "${library}_${start_offset}_${end_offset}" + // bamadap/picodup aren't on bioconda yet, so CI can't install real binaries; -stub-run + // instead replays real output this exact chunk produced on a genuine (local-machine) + // run of the real pipeline against these same fixtures, captured once under + // tests/fixtures/stub_outputs/. Regenerate those fixtures if trim_and_align.nf's + // logic, its conda pins, or the test fixtures themselves change. + def fixture_dir = "${workflow.projectDir}/tests/fixtures/stub_outputs/trim_and_align" + """ + cp ${fixture_dir}/${chunk}.aln.bam . + cp ${fixture_dir}/${chunk}.fastp.json . + cp ${fixture_dir}/${chunk}.nonconverted_counts.tsv . + touch ${library}.log.bwamem + """ +} diff --git a/nextflow.config b/nextflow.config index b83c626..5b140d9 100644 --- a/nextflow.config +++ b/nextflow.config @@ -26,8 +26,20 @@ params { // null means no cap — per-process defaults in conf/base.config apply. max_memory = null + // Optional cpu cap applied to labeled processes (e.g. --max_cpus 8). + // null means no cap — per-process defaults in conf/base.config apply. + max_cpus = null + + // Binaries not yet packaged for bioconda; deployed to production via Capistrano + // (nebiolabs/bamadap, nebiolabs/picodup). Override with a different absolute path + // (e.g. conf/test.config) until bioconda packaging lands. + bamadap_bin = '/mnt/bioinfo/prg/bamadap/current/bin/bamadap' + picodup_bin = '/mnt/bioinfo/prg/picodup/current/bin/picodup' + + picodup_optical_model = 'ellipse' + picodup_optical_distance = null // null => derive from instrument name, as Picard path does + // NEB only - path_to_ngs_agg = null workflow_name_modifier = null } @@ -85,12 +97,6 @@ env { PYTHONNOUSERSITE = 1 } -executor { - name = 'sge' - queueSize = 10000 - jobName = { "${task.process.tokenize(':')[-1]}" } -} - process.shell = ['/bin/bash', '-euo', 'pipefail'] timeline { diff --git a/tests/fastq_to_ubam.nf.test.snap b/tests/fastq_to_ubam.nf.test.snap index ba86327..545084b 100644 --- a/tests/fastq_to_ubam.nf.test.snap +++ b/tests/fastq_to_ubam.nf.test.snap @@ -2,9 +2,9 @@ "fastq to uBam workflow - single-end": { "content": [ { - "tasksCount": 2, + "tasksCount": 4, "tasksFailed": 0, - "tasksSucceeded": 2 + "tasksSucceeded": 4 }, [ "ubam", @@ -19,18 +19,18 @@ } ] ], - "timestamp": "2026-07-29T14:20:52.691934926", + "timestamp": "2026-09-07T22:24:27.574255349", "meta": { - "nf-test": "0.9.5", - "nextflow": "26.04.0" + "nf-test": "0.9.4", + "nextflow": "25.10.4" } }, "fastq to uBam workflow - non-standard SRA headers (issue #15)": { "content": [ { - "tasksCount": 1, + "tasksCount": 2, "tasksFailed": 0, - "tasksSucceeded": 1 + "tasksSucceeded": 2 }, [ "ubam", @@ -45,18 +45,18 @@ } ] ], - "timestamp": "2026-07-29T14:20:59.43877386", + "timestamp": "2026-09-07T22:24:36.675298676", "meta": { - "nf-test": "0.9.5", - "nextflow": "26.04.0" + "nf-test": "0.9.4", + "nextflow": "25.10.4" } }, "fastq to uBam workflow - paired-end": { "content": [ { - "tasksCount": 2, + "tasksCount": 4, "tasksFailed": 0, - "tasksSucceeded": 2 + "tasksSucceeded": 4 }, [ "ubam", @@ -71,10 +71,10 @@ } ] ], - "timestamp": "2026-07-29T14:20:45.730120318", + "timestamp": "2026-09-07T22:24:18.57790468", "meta": { - "nf-test": "0.9.5", - "nextflow": "26.04.0" + "nf-test": "0.9.4", + "nextflow": "25.10.4" } } } \ No newline at end of file diff --git a/tests/fixtures/reference_files/reference.dict b/tests/fixtures/reference_files/reference.dict new file mode 100644 index 0000000..918b1b0 --- /dev/null +++ b/tests/fixtures/reference_files/reference.dict @@ -0,0 +1,2 @@ +@HD VN:1.0 SO:unsorted +@SQ SN:chr1 LN:284940 M5:e2d98a7cc072e12f74f7ef08529c1cba UR:file:///appdev/langhorst/src/EM-seq/tests/fixtures/reference_files/reference.fa diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.ds.1.markdups_log b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.ds.1.markdups_log new file mode 100644 index 0000000..10ff3b3 --- /dev/null +++ b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.ds.1.markdups_log @@ -0,0 +1,10 @@ +## htsjdk.samtools.metrics.StringHeader +# /appdev/langhorst/src/picodup/target-el8/release/picodup --input /dev/stdin --output emseq-test1.ds.1.md.bam --metrics emseq-test1.ds.1.markdups_log --threads 14 --optical-model ellipse --optical-distance 2500 --tagging-policy All +## htsjdk.samtools.metrics.StringHeader +# Started on: Mon Sep 07 22:25:49 EDT 2026 + +## METRICS CLASS picard.sam.DuplicationMetrics +LIBRARY UNPAIRED_READS_EXAMINED READ_PAIRS_EXAMINED SECONDARY_OR_SUPPLEMENTARY_RDS UNMAPPED_READS UNPAIRED_READ_DUPLICATES READ_PAIR_DUPLICATES READ_PAIR_OPTICAL_DUPLICATES PERCENT_DUPLICATION ESTIMATED_LIBRARY_SIZE +emseq-test1.ds.1 745 0 18 0 14 0 0 0.018792 + + diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.ds.1.md.bai b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.ds.1.md.bai new file mode 100644 index 0000000..68ffd4f Binary files /dev/null and b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.ds.1.md.bai differ diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.ds.1.md.bam b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.ds.1.md.bam new file mode 100644 index 0000000..b528f79 Binary files /dev/null and b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.ds.1.md.bam differ diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.markdups_log b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.markdups_log new file mode 100644 index 0000000..507e936 --- /dev/null +++ b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.markdups_log @@ -0,0 +1,13 @@ +## htsjdk.samtools.metrics.StringHeader +# /appdev/langhorst/src/picodup/target-el8/release/picodup --input /dev/stdin --output emseq-test1.md.bam --metrics emseq-test1.markdups_log --threads 14 --optical-model ellipse --optical-distance 100 --tagging-policy All +## htsjdk.samtools.metrics.StringHeader +# Started on: Mon Sep 07 22:25:29 EDT 2026 + +## METRICS CLASS picard.sam.DuplicationMetrics +LIBRARY UNPAIRED_READS_EXAMINED READ_PAIRS_EXAMINED SECONDARY_OR_SUPPLEMENTARY_RDS UNMAPPED_READS UNPAIRED_READ_DUPLICATES READ_PAIR_DUPLICATES READ_PAIR_OPTICAL_DUPLICATES PERCENT_DUPLICATION ESTIMATED_LIBRARY_SIZE +Oct25_PV_10ngR3_F6 264 635 1 264 24 0 0 0.015645 + +## HISTOGRAM java.lang.Double +BIN CoverageMult all_sets optical_sets non_optical_sets +1.0 0.000000 623 0 623 + diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.md.bai b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.md.bai new file mode 100644 index 0000000..cc29ca9 Binary files /dev/null and b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.md.bai differ diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.md.bam b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.md.bam new file mode 100644 index 0000000..de14fcb Binary files /dev/null and b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test1.md.bam differ diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.ds.1.markdups_log b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.ds.1.markdups_log new file mode 100644 index 0000000..8b1b551 --- /dev/null +++ b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.ds.1.markdups_log @@ -0,0 +1,10 @@ +## htsjdk.samtools.metrics.StringHeader +# /appdev/langhorst/src/picodup/target-el8/release/picodup --input /dev/stdin --output emseq-test2.ds.1.md.bam --metrics emseq-test2.ds.1.markdups_log --threads 14 --optical-model ellipse --optical-distance 2500 --tagging-policy All +## htsjdk.samtools.metrics.StringHeader +# Started on: Mon Sep 07 22:25:49 EDT 2026 + +## METRICS CLASS picard.sam.DuplicationMetrics +LIBRARY UNPAIRED_READS_EXAMINED READ_PAIRS_EXAMINED SECONDARY_OR_SUPPLEMENTARY_RDS UNMAPPED_READS UNPAIRED_READ_DUPLICATES READ_PAIR_DUPLICATES READ_PAIR_OPTICAL_DUPLICATES PERCENT_DUPLICATION ESTIMATED_LIBRARY_SIZE +emseq-test2.ds.1 48 0 0 0 0 0 0 0.000000 + + diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.ds.1.md.bai b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.ds.1.md.bai new file mode 100644 index 0000000..f094e5f Binary files /dev/null and b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.ds.1.md.bai differ diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.ds.1.md.bam b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.ds.1.md.bam new file mode 100644 index 0000000..70de194 Binary files /dev/null and b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.ds.1.md.bam differ diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.markdups_log b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.markdups_log new file mode 100644 index 0000000..e6b775e --- /dev/null +++ b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.markdups_log @@ -0,0 +1,13 @@ +## htsjdk.samtools.metrics.StringHeader +# /appdev/langhorst/src/picodup/target-el8/release/picodup --input /dev/stdin --output emseq-test2.md.bam --metrics emseq-test2.markdups_log --threads 14 --optical-model ellipse --optical-distance 100 --tagging-policy All +## htsjdk.samtools.metrics.StringHeader +# Started on: Mon Sep 07 22:25:29 EDT 2026 + +## METRICS CLASS picard.sam.DuplicationMetrics +LIBRARY UNPAIRED_READS_EXAMINED READ_PAIRS_EXAMINED SECONDARY_OR_SUPPLEMENTARY_RDS UNMAPPED_READS UNPAIRED_READ_DUPLICATES READ_PAIR_DUPLICATES READ_PAIR_OPTICAL_DUPLICATES PERCENT_DUPLICATION ESTIMATED_LIBRARY_SIZE +Oct25_PV_10ngR3_F6 264 635 1 264 23 0 0 0.014993 + +## HISTOGRAM java.lang.Double +BIN CoverageMult all_sets optical_sets non_optical_sets +1.0 0.000000 624 0 624 + diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.md.bai b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.md.bai new file mode 100644 index 0000000..0fb78c3 Binary files /dev/null and b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.md.bai differ diff --git a/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.md.bam b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.md.bam new file mode 100644 index 0000000..a5dc419 Binary files /dev/null and b/tests/fixtures/stub_outputs/merge_and_picodup/emseq-test2.md.bam differ diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test1.ds.1_0_257200.aln.bam b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1.ds.1_0_257200.aln.bam new file mode 100644 index 0000000..f84c365 Binary files /dev/null and b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1.ds.1_0_257200.aln.bam differ diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test1.ds.1_0_257200.fastp.json b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1.ds.1_0_257200.fastp.json new file mode 100644 index 0000000..397f181 --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1.ds.1_0_257200.fastp.json @@ -0,0 +1,5510 @@ +{ + "summary": { + "bamadap_version": "0.1.0", + "fastp_version": "0.1.0", + "sequencing": "single end (150 cycles)", + "before_filtering": { + "total_reads": 2500, + "total_bases": 375000, + "q20_bases": 368284, + "q30_bases": 356222, + "q20_rate": 0.9820906666666667, + "q30_rate": 0.9499253333333333, + "read1_mean_length": 150, + "gc_content": 0.21490933333333334 + }, + "after_filtering": { + "total_reads": 2500, + "total_bases": 371387, + "q20_bases": 364731, + "q30_bases": 352753, + "q20_rate": 0.9820779941139566, + "q30_rate": 0.9498259228244392, + "read1_mean_length": 148, + "gc_content": 0.21198103326179968 + } + }, + "filtering_result": { + "passed_filter_reads": 2500, + "low_quality_reads": 0, + "too_many_N_reads": 0, + "adapter_dimer_reads": 0, + "too_short_reads": 0, + "too_long_reads": 0, + "low_complexity_reads": 0 + }, + "duplication": { + "rate": 0.0 + }, + "adapter_cutting": { + "adapter_trimmed_reads": 156, + "adapter_trimmed_bases": 3613, + "read1_adapter_sequence": "AGATCGGAAGAGC", + "read2_adapter_sequence": "AGATCGGAAGAGC", + "read1_adapter_counts": { + "AGATC": 6, + "AGATCG": 7, + "AGATCGG": 6, + "AGATCGGA": 4, + "AGATCGGAA": 6, + "AGATCGGAAG": 7, + "AGATCGGAAGA": 7, + "AGATCGGAAGAG": 7, 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0000000..4b1db9b --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1.ds.1_0_257200.nonconverted_counts.tsv @@ -0,0 +1 @@ +chr1 0 diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_0_250000.aln.bam b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_0_250000.aln.bam new file mode 100644 index 0000000..0f4388e Binary files /dev/null and b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_0_250000.aln.bam differ diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_0_250000.fastp.json b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_0_250000.fastp.json new file mode 100644 index 0000000..0c4f080 --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_0_250000.fastp.json @@ -0,0 +1,8219 @@ +{ + "summary": { + "bamadap_version": "0.1.0", + "fastp_version": "0.1.0", + "sequencing": "paired end (76 cycles + 76 cycles)", + "before_filtering": { + "total_reads": 5680, + "total_bases": 431680, + 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"GGGGA": 0, + "GGGGT": 0, + "GGGGC": 0, + "GGGGG": 0 + }, + "overrepresented_sequences": {} + }, + "command": "/appdev/langhorst/src/bamadap/target-el8/release/bamadap --input /tmp/emseq-test1_0_250000.slice.bam --format fastq --no-mate-suffix -l 2 --disable-quality-filtering --adapter-fasta illumina_adapters.fa --trim-poly-g --threads 4 --json emseq-test1_0_250000.fastp.json" +} \ No newline at end of file diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_0_250000.nonconverted_counts.tsv b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_0_250000.nonconverted_counts.tsv new file mode 100644 index 0000000..4b1db9b --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_0_250000.nonconverted_counts.tsv @@ -0,0 +1 @@ +chr1 0 diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_250000_421605.aln.bam b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_250000_421605.aln.bam new file mode 100644 index 0000000..734fe90 Binary files /dev/null and b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_250000_421605.aln.bam differ diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_250000_421605.fastp.json b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_250000_421605.fastp.json new file mode 100644 index 0000000..4fcaf3b --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_250000_421605.fastp.json @@ -0,0 +1,8205 @@ +{ + "summary": { + "bamadap_version": "0.1.0", + "fastp_version": "0.1.0", + "sequencing": "paired end (76 cycles + 76 cycles)", + "before_filtering": { + "total_reads": 3854, + "total_bases": 292904, + "q20_bases": 275601, + "q30_bases": 268156, + "q20_rate": 0.9409260371999016, + "q30_rate": 0.9155081528418868, + "read1_mean_length": 76, + "read2_mean_length": 76, + "gc_content": 0.21639513287630077 + }, + "after_filtering": { + "total_reads": 3848, + "total_bases": 290833, + "q20_bases": 273935, + "q30_bases": 266561, + "q20_rate": 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b/tests/fixtures/stub_outputs/trim_and_align/emseq-test1_250000_421605.nonconverted_counts.tsv @@ -0,0 +1 @@ +chr1 0 diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test2.ds.1_0_16821.aln.bam b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2.ds.1_0_16821.aln.bam new file mode 100644 index 0000000..ac12614 Binary files /dev/null and b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2.ds.1_0_16821.aln.bam differ diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test2.ds.1_0_16821.fastp.json b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2.ds.1_0_16821.fastp.json new file mode 100644 index 0000000..9719ab6 --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2.ds.1_0_16821.fastp.json @@ -0,0 +1,4387 @@ +{ + "summary": { + "bamadap_version": "0.1.0", + "fastp_version": "0.1.0", + "sequencing": "single end (99 cycles)", + "before_filtering": { + "total_reads": 369, + "total_bases": 36531, + "q20_bases": 36142, + "q30_bases": 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a/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_0_250000.aln.bam b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_0_250000.aln.bam new file mode 100644 index 0000000..7d2de7a Binary files /dev/null and b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_0_250000.aln.bam differ diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_0_250000.fastp.json b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_0_250000.fastp.json new file mode 100644 index 0000000..27b66cf --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_0_250000.fastp.json @@ -0,0 +1,8219 @@ +{ + "summary": { + "bamadap_version": "0.1.0", + "fastp_version": "0.1.0", + "sequencing": "paired end (76 cycles + 76 cycles)", + "before_filtering": { + "total_reads": 5680, + "total_bases": 431680, + "q20_bases": 405386, + "q30_bases": 393968, + "q20_rate": 0.9390891401037805, + "q30_rate": 0.9126389918458118, + "read1_mean_length": 76, + "read2_mean_length": 76, + 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{} + }, + "command": "/appdev/langhorst/src/bamadap/target-el8/release/bamadap --input /tmp/emseq-test2_0_250000.slice.bam --format fastq --no-mate-suffix -l 2 --disable-quality-filtering --adapter-fasta illumina_adapters.fa --trim-poly-g --threads 4 --json emseq-test2_0_250000.fastp.json" +} \ No newline at end of file diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_0_250000.nonconverted_counts.tsv b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_0_250000.nonconverted_counts.tsv new file mode 100644 index 0000000..4b1db9b --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_0_250000.nonconverted_counts.tsv @@ -0,0 +1 @@ +chr1 0 diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_250000_426231.aln.bam b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_250000_426231.aln.bam new file mode 100644 index 0000000..7d73370 Binary files /dev/null and b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_250000_426231.aln.bam differ diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_250000_426231.fastp.json b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_250000_426231.fastp.json new file mode 100644 index 0000000..a0fe1cd --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_250000_426231.fastp.json @@ -0,0 +1,8205 @@ +{ + "summary": { + "bamadap_version": "0.1.0", + "fastp_version": "0.1.0", + "sequencing": "paired end (76 cycles + 76 cycles)", + "before_filtering": { + "total_reads": 3852, + "total_bases": 292752, + "q20_bases": 275514, + "q30_bases": 268079, + "q20_rate": 0.9411173962944746, + "q30_rate": 0.9157204733016342, + "read1_mean_length": 76, + "read2_mean_length": 76, + "gc_content": 0.21632986281904137 + }, + "after_filtering": { + "total_reads": 3846, + "total_bases": 290681, + "q20_bases": 273848, + "q30_bases": 266484, + "q20_rate": 0.9420911583488429, + "q30_rate": 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"GGCAC": 0, + "GGCAG": 0, + "GGCTA": 0, + "GGCTT": 0, + "GGCTC": 0, + "GGCTG": 0, + "GGCCA": 0, + "GGCCT": 0, + "GGCCC": 0, + "GGCCG": 0, + "GGCGA": 0, + "GGCGT": 0, + "GGCGC": 0, + "GGCGG": 0, + "GGGAA": 0, + "GGGAT": 0, + "GGGAC": 0, + "GGGAG": 0, + "GGGTA": 0, + "GGGTT": 0, + "GGGTC": 0, + "GGGTG": 0, + "GGGCA": 0, + "GGGCT": 0, + "GGGCC": 0, + "GGGCG": 0, + "GGGGA": 0, + "GGGGT": 0, + "GGGGC": 0, + "GGGGG": 0 + }, + "overrepresented_sequences": {} + }, + "command": "/appdev/langhorst/src/bamadap/target-el8/release/bamadap --input /tmp/emseq-test2_250000_426231.slice.bam --format fastq --no-mate-suffix -l 2 --disable-quality-filtering --adapter-fasta illumina_adapters.fa --trim-poly-g --threads 4 --json emseq-test2_250000_426231.fastp.json" +} \ No newline at end of file diff --git a/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_250000_426231.nonconverted_counts.tsv b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_250000_426231.nonconverted_counts.tsv new file mode 100644 index 0000000..4b1db9b --- /dev/null +++ b/tests/fixtures/stub_outputs/trim_and_align/emseq-test2_250000_426231.nonconverted_counts.tsv @@ -0,0 +1 @@ +chr1 0 diff --git a/tests/main.nf.test b/tests/main.nf.test index 4690a8c..67ac147 100644 --- a/tests/main.nf.test +++ b/tests/main.nf.test @@ -74,11 +74,11 @@ nextflow_pipeline { test("EM-seq workflow uBam - single-end") { when { - params.ubam_dir = "$projectDir/tests/fixtures/ubam/" + params.ubam_dir = "$projectDir/tests/fixtures/ubam_single_end/" params.genome = 'test' params.email = 'foo@example.com' params.flowcell = "test_pipeline" - params.bamslice_chunk_size = 250000 + params.bamslice_chunk_size = 300000 params.enable_neb_agg = true params.single_end = true params.adapter_set = "test" @@ -86,33 +86,31 @@ nextflow_pipeline { } then { - def bwameth_file_1 = bam("${launchDir}/test_output/bwameth_align/emseq-test1_0_250000.aln.bam").getStatistics() - def bwameth_file_2 = bam("${launchDir}/test_output/bwameth_align/emseq-test1_250000_421605.aln.bam").getStatistics() - def fastqc = path("${launchDir}/test_output/stats/fastqc/emseq-test1.md_fastqc.html").exists() - def fastp = path("${launchDir}/test_output/fastp/emseq-test1.fastp.json").text.tokenize('\n')[1..33] - def markdup = bam("${launchDir}/test_output/markduped_bams/emseq-test1.md.bam").getStatistics() - def markdup_log = path("${launchDir}/test_output/stats/markdups/emseq-test1.markdups_log").text.tokenize('\n')[5..6] - def idxstats = path("${launchDir}/test_output/stats/idxstats/emseq-test1.idxstat").text.tokenize('\n') - def flagstats = path("${launchDir}/test_output/stats/flagstats/emseq-test1.flagstat").text.tokenize('\n') - def gc_bias = path("${launchDir}/test_output/stats/gc_bias/emseq-test1.gc_metrics").text.tokenize('\n')[5..-1] - def alignment_metrics = path("${launchDir}/test_output/stats/picard_alignment_metrics/emseq-test1.alignment_summary_metrics.txt").text.tokenize('\n')[5..8] - def methyldackel_extract = path("${launchDir}/test_output/methylDackelExtracts/emseq-test1_CpG.methylKit.gz").md5 - def mbias = path("${launchDir}/test_output/methylDackelExtracts/mbias/emseq-test1.combined_mbias.tsv").md5 - def nonconverted = path("${launchDir}/test_output/bwameth_align/emseq-test1_0_250000.nonconverted_counts.tsv").text.tokenize('\n') - def nonconverted_combined = path("${launchDir}/test_output/stats/nonconverted_counts/emseq-test1.nonconverted_counts.for_agg.tsv").text.tokenize('\n') + def bwameth_file_1 = bam("${launchDir}/test_output/bwameth_align/emseq-test1.ds.1_0_257200.aln.bam").getStatistics() + def fastqc = path("${launchDir}/test_output/stats/fastqc/emseq-test1.ds.1.md_fastqc.html").exists() + def fastp = path("${launchDir}/test_output/fastp/emseq-test1.ds.1.fastp.json").text.tokenize('\n')[1..33] + def markdup = bam("${launchDir}/test_output/markduped_bams/emseq-test1.ds.1.md.bam").getStatistics() + def markdup_log = path("${launchDir}/test_output/stats/markdups/emseq-test1.ds.1.markdups_log").text.tokenize('\n')[5..6] + def idxstats = path("${launchDir}/test_output/stats/idxstats/emseq-test1.ds.1.idxstat").text.tokenize('\n') + def flagstats = path("${launchDir}/test_output/stats/flagstats/emseq-test1.ds.1.flagstat").text.tokenize('\n') + def gc_bias = path("${launchDir}/test_output/stats/gc_bias/emseq-test1.ds.1.gc_metrics").text.tokenize('\n')[5..-1] + def alignment_metrics = path("${launchDir}/test_output/stats/picard_alignment_metrics/emseq-test1.ds.1.alignment_summary_metrics.txt").text.tokenize('\n')[5..8] + def methyldackel_extract = path("${launchDir}/test_output/methylDackelExtracts/emseq-test1.ds.1_CpG.methylKit.gz").md5 + def mbias = path("${launchDir}/test_output/methylDackelExtracts/mbias/emseq-test1.ds.1.combined_mbias.tsv").md5 + def nonconverted = path("${launchDir}/test_output/bwameth_align/emseq-test1.ds.1_0_257200.nonconverted_counts.tsv").text.tokenize('\n') + def nonconverted_combined = path("${launchDir}/test_output/stats/nonconverted_counts/emseq-test1.ds.1.nonconverted_counts.for_agg.tsv").text.tokenize('\n') def combined_summaries = path("${launchDir}/test_output/methylKit_intersections/positional_summaries_combined.tsv").exists() - def positional_summary = path("${launchDir}/test_output/methylKit_intersections/emseq-test1_vs_emseq_test_regions_positional_summary.tsv").text.tokenize('\n') - def methylkit_intersections = path("${launchDir}/test_output/methylKit_intersections/emseq-test2_vs_emseq_test_regions_intersections.tsv").text.tokenize('\n') - def region_summary = path("${launchDir}/test_output/methylKit_intersections/emseq-test2_vs_emseq_test_regions_region_summary.tsv").text.tokenize('\n') - def tasmanian = path("${launchDir}/test_output/stats/tasmanian/emseq-test1.tasmanian.csv").exists() + def positional_summary = path("${launchDir}/test_output/methylKit_intersections/emseq-test1.ds.1_vs_emseq_test_regions_positional_summary.tsv").text.tokenize('\n') + def methylkit_intersections = path("${launchDir}/test_output/methylKit_intersections/emseq-test2.ds.1_vs_emseq_test_regions_intersections.tsv").text.tokenize('\n') + def region_summary = path("${launchDir}/test_output/methylKit_intersections/emseq-test2.ds.1_vs_emseq_test_regions_region_summary.tsv").text.tokenize('\n') + def tasmanian = path("${launchDir}/test_output/stats/tasmanian/emseq-test1.ds.1.tasmanian.csv").exists() def multiqc = path("${launchDir}/test_output/EM-seq-Alignment-Summary-test_pipeline_multiqc_report.html").exists() - def ngs_agg = path("${launchDir}/test_output/aggregate_results/emseq-test1.arguments.txt").text.tokenize('\n') + def ngs_agg = path("${launchDir}/test_output/aggregate_results/emseq-test1.ds.1.arguments.txt").text.tokenize('\n') assertAll( { assert workflow.success }, { assert snapshot(workflow.trace, ["bwameth_file_1", bwameth_file_1], - ["bwameth_file_2", bwameth_file_2], ["fastqc", fastqc], ["fastp", fastp], ["markdup", markdup], diff --git a/tests/main.nf.test.snap b/tests/main.nf.test.snap index fbf1058..c199cde 100644 --- a/tests/main.nf.test.snap +++ b/tests/main.nf.test.snap @@ -2,32 +2,20 @@ "EM-seq workflow uBam - single-end": { "content": [ { - "tasksCount": 47, + "tasksCount": 41, "tasksFailed": 0, - "tasksSucceeded": 47 + "tasksSucceeded": 41 }, [ "bwameth_file_1", { - "maxReadLength": 76, - "minReadLength": 44, - "meanReadLength": 75, - "maxQuality": 60, - "minQuality": 0, - "meanQuality": 10, - "readCount": 702 - } - ], - [ - "bwameth_file_2", - { - "maxReadLength": 76, - "minReadLength": 52, - "meanReadLength": 75, + "maxReadLength": 150, + "minReadLength": 53, + "meanReadLength": 146, "maxQuality": 60, "minQuality": 0, - "meanQuality": 8, - "readCount": 399 + "meanQuality": 41, + "readCount": 763 } ], [ @@ -38,35 +26,35 @@ "fastp", [ "\t\"summary\": {", - "\t\t\"fastp_version\": \"1.3.6\",", - "\t\t\"sequencing\": \"single end (76 cycles)\",", + "\t\t\"fastp_version\": \"0.1.0\",", + "\t\t\"sequencing\": \"single end (150 cycles)\",", "\t\t\"before_filtering\": {", - "\t\t\t\"total_reads\": 9534,", - "\t\t\t\"total_bases\": 724584,", - "\t\t\t\"q20_bases\": 681052,", - "\t\t\t\"q30_bases\": 662196,", - "\t\t\t\"q20_rate\": 0.939921,", - "\t\t\t\"q30_rate\": 0.913898,", - "\t\t\t\"read1_mean_length\": 76,", - "\t\t\t\"gc_content\": 0.218667", + "\t\t\t\"total_reads\": 2500,", + "\t\t\t\"total_bases\": 375000,", + "\t\t\t\"q20_bases\": 368284,", + "\t\t\t\"q30_bases\": 356222,", + "\t\t\t\"q20_rate\": 0.982091,", + "\t\t\t\"q30_rate\": 0.949925,", + "\t\t\t\"read1_mean_length\": 150,", + "\t\t\t\"gc_content\": 0.214909", "\t\t},", "\t\t\"after_filtering\": {", - "\t\t\t\"total_reads\": 9530,", - "\t\t\t\"total_bases\": 721079,", - "\t\t\t\"q20_bases\": 678095,", - "\t\t\t\"q30_bases\": 659406,", - "\t\t\t\"q20_rate\": 0.940389,", - "\t\t\t\"q30_rate\": 0.914471,", - "\t\t\t\"read1_mean_length\": 75,", - "\t\t\t\"gc_content\": 0.216831", + "\t\t\t\"total_reads\": 2500,", + "\t\t\t\"total_bases\": 371387,", + "\t\t\t\"q20_bases\": 364731,", + "\t\t\t\"q30_bases\": 352753,", + "\t\t\t\"q20_rate\": 0.982078,", + "\t\t\t\"q30_rate\": 0.949826,", + "\t\t\t\"read1_mean_length\": 148,", + "\t\t\t\"gc_content\": 0.211981", "\t\t}", "\t},", "\t\"filtering_result\": {", - "\t\t\"passed_filter_reads\": 9530,", + "\t\t\"passed_filter_reads\": 2500,", "\t\t\"low_quality_reads\": 0,", "\t\t\"too_many_N_reads\": 0,", "\t\t\"adapter_dimer_reads\": 0,", - "\t\t\"too_short_reads\": 4,", + "\t\t\"too_short_reads\": 0,", "\t\t\"too_long_reads\": 0", "\t},", "\t\"duplication\": {" @@ -75,40 +63,40 @@ [ "markdup", { - "maxReadLength": 76, - "minReadLength": 44, - "meanReadLength": 75, + "maxReadLength": 150, + "minReadLength": 53, + "meanReadLength": 146, "maxQuality": 60, "minQuality": 0, - "meanQuality": 9, - "readCount": 1101 + "meanQuality": 41, + "readCount": 763 } ], [ "markdup_log", [ "LIBRARY\tUNPAIRED_READS_EXAMINED\tREAD_PAIRS_EXAMINED\tSECONDARY_OR_SUPPLEMENTARY_RDS\tUNMAPPED_READS\tUNPAIRED_READ_DUPLICATES\tREAD_PAIR_DUPLICATES\tREAD_PAIR_OPTICAL_DUPLICATES\tPERCENT_DUPLICATION\tESTIMATED_LIBRARY_SIZE", - "Oct25_PV_10ngR3_F6\t1100\t0\t1\t0\t81\t0\t0\t0.073636\t" + "emseq-test1.ds.1\t745\t0\t18\t0\t14\t0\t0\t0.018792\t" ] ], [ "idxstats", [ - "chr1\t284940\t1101\t0", + "chr1\t284940\t763\t0", "*\t0\t0\t0" ] ], [ "flagstats", [ - "1095 + 6 in total (QC-passed reads + QC-failed reads)", - "1094 + 6 primary", - "1 + 0 secondary", + "763 + 0 in total (QC-passed reads + QC-failed reads)", + "745 + 0 primary", + "18 + 0 secondary", "0 + 0 supplementary", - "81 + 0 duplicates", - "81 + 0 primary duplicates", - "1095 + 6 mapped (100.00% : 100.00%)", - "1094 + 6 primary mapped (100.00% : 100.00%)", + "14 + 0 duplicates", + "14 + 0 primary duplicates", + "763 + 0 mapped (100.00% : N/A)", + "745 + 0 primary mapped (100.00% : N/A)", "0 + 0 paired in sequencing", "0 + 0 read1", "0 + 0 read2", @@ -123,59 +111,59 @@ "gc_bias", [ "ACCUMULATION_LEVEL\tREADS_USED\tGC\tWINDOWS\tREAD_STARTS\tMEAN_BASE_QUALITY\tNORMALIZED_COVERAGE\tERROR_BAR_WIDTH\tSAMPLE\tLIBRARY\tREAD_GROUP", - "All Reads\tALL\t0\t17252\t122\t6\t1.8295\t0.165635\t\t\t", - "All Reads\tALL\t1\t3503\t37\t6\t2.732585\t0.449234\t\t\t", - "All Reads\tALL\t2\t5640\t47\t6\t2.155911\t0.314472\t\t\t", - "All Reads\tALL\t3\t3475\t40\t6\t2.977949\t0.470855\t\t\t", - "All Reads\tALL\t4\t4275\t45\t5\t2.723256\t0.405959\t\t\t", - "All Reads\tALL\t5\t3480\t18\t5\t1.338152\t0.315405\t\t\t", - "All Reads\tALL\t6\t3400\t23\t5\t1.750093\t0.36492\t\t\t", - "All Reads\tALL\t7\t3235\t19\t6\t1.519468\t0.34859\t\t\t", - "All Reads\tALL\t8\t3640\t7\t6\t0.497518\t0.188044\t\t\t", - "All Reads\tALL\t9\t3412\t11\t6\t0.834057\t0.251478\t\t\t", - "All Reads\tALL\t10\t3608\t12\t6\t0.860452\t0.248391\t\t\t", - "All Reads\tALL\t11\t3699\t12\t5\t0.839284\t0.24228\t\t\t", - "All Reads\tALL\t12\t3840\t7\t6\t0.471606\t0.17825\t\t\t", - "All Reads\tALL\t13\t3874\t8\t7\t0.534248\t0.188885\t\t\t", - "All Reads\tALL\t14\t3693\t15\t6\t1.05081\t0.271318\t\t\t", - "All Reads\tALL\t15\t3986\t22\t7\t1.427899\t0.304429\t\t\t", - "All Reads\tALL\t16\t3911\t17\t7\t1.124536\t0.27274\t\t\t", - "All Reads\tALL\t17\t3778\t16\t7\t1.095646\t0.273911\t\t\t", - "All Reads\tALL\t18\t3793\t25\t7\t1.705176\t0.341035\t\t\t", - "All Reads\tALL\t19\t3698\t30\t7\t2.098778\t0.383183\t\t\t", - "All Reads\tALL\t20\t3735\t28\t7\t1.939454\t0.366522\t\t\t", - "All Reads\tALL\t21\t3759\t34\t5\t2.340015\t0.401309\t\t\t", - "All Reads\tALL\t22\t3461\t35\t6\t2.616246\t0.442226\t\t\t", - "All Reads\tALL\t23\t3475\t41\t7\t3.052398\t0.476705\t\t\t", - "All Reads\tALL\t24\t3393\t62\t7\t4.727374\t0.600377\t\t\t", - "All Reads\tALL\t25\t3365\t52\t7\t3.997886\t0.554407\t\t\t", - "All Reads\tALL\t26\t3200\t43\t6\t3.476407\t0.530147\t\t\t", - "All Reads\tALL\t27\t2988\t38\t8\t3.290146\t0.533732\t\t\t", - "All Reads\tALL\t28\t2852\t35\t6\t3.174904\t0.536657\t\t\t", - "All Reads\tALL\t29\t2805\t29\t8\t2.674713\t0.496682\t\t\t", - "All Reads\tALL\t30\t2833\t10\t5\t0.913199\t0.288779\t\t\t", - "All Reads\tALL\t31\t2731\t23\t7\t2.178805\t0.454312\t\t\t", - "All Reads\tALL\t32\t2760\t25\t8\t2.343382\t0.468676\t\t\t", - "All Reads\tALL\t33\t2776\t16\t6\t1.49112\t0.37278\t\t\t", - "All Reads\tALL\t34\t2475\t15\t5\t1.567935\t0.404839\t\t\t", - "All Reads\tALL\t35\t2672\t14\t6\t1.355513\t0.362276\t\t\t", - "All Reads\tALL\t36\t2796\t7\t8\t0.647699\t0.244807\t\t\t", - "All Reads\tALL\t37\t2850\t9\t11\t0.816977\t0.272326\t\t\t", - "All Reads\tALL\t38\t2826\t11\t7\t1.007007\t0.303624\t\t\t", - "All Reads\tALL\t39\t3054\t4\t10\t0.338847\t0.169423\t\t\t", - "All Reads\tALL\t40\t3449\t6\t8\t0.45006\t0.183736\t\t\t", - "All Reads\tALL\t41\t4012\t9\t5\t0.580355\t0.193452\t\t\t", - "All Reads\tALL\t42\t4971\t7\t5\t0.364306\t0.137695\t\t\t", - "All Reads\tALL\t43\t5630\t5\t6\t0.22976\t0.102752\t\t\t", - "All Reads\tALL\t44\t6409\t2\t11\t0.080733\t0.057087\t\t\t", - "All Reads\tALL\t45\t7297\t2\t8\t0.070908\t0.05014\t\t\t", - "All Reads\tALL\t46\t8146\t2\t10\t0.063518\t0.044914\t\t\t", - "All Reads\tALL\t47\t8876\t1\t8\t0.029147\t0.029147\t\t\t", + "All Reads\tALL\t0\t17252\t5\t8\t0.108195\t0.048386\t\t\t", + "All Reads\tALL\t1\t3503\t2\t10\t0.21314\t0.150713\t\t\t", + "All Reads\tALL\t2\t5640\t2\t9\t0.132381\t0.093608\t\t\t", + "All Reads\tALL\t3\t3475\t0\t0\t0\t0\t\t\t", + "All Reads\tALL\t4\t4275\t2\t12\t0.17465\t0.123496\t\t\t", + "All Reads\tALL\t5\t3480\t2\t0\t0.214549\t0.151709\t\t\t", + "All Reads\tALL\t6\t3400\t1\t0\t0.109798\t0.109798\t\t\t", + "All Reads\tALL\t7\t3235\t3\t11\t0.346196\t0.199876\t\t\t", + "All Reads\tALL\t8\t3640\t7\t30\t0.717913\t0.271345\t\t\t", + "All Reads\tALL\t9\t3412\t6\t17\t0.656473\t0.268004\t\t\t", + "All Reads\tALL\t10\t3608\t4\t28\t0.413874\t0.206937\t\t\t", + "All Reads\tALL\t11\t3699\t15\t33\t1.513847\t0.390873\t\t\t", + "All Reads\tALL\t12\t3840\t20\t20\t1.944347\t0.434769\t\t\t", + "All Reads\tALL\t13\t3874\t25\t17\t2.409103\t0.481821\t\t\t", + "All Reads\tALL\t14\t3693\t23\t16\t2.325003\t0.484797\t\t\t", + "All Reads\tALL\t15\t3986\t38\t19\t3.558945\t0.577337\t\t\t", + "All Reads\tALL\t16\t3911\t25\t14\t2.386311\t0.477262\t\t\t", + "All Reads\tALL\t17\t3778\t18\t14\t1.778629\t0.419227\t\t\t", + "All Reads\tALL\t18\t3793\t36\t16\t3.543191\t0.590532\t\t\t", + "All Reads\tALL\t19\t3698\t38\t16\t3.836115\t0.6223\t\t\t", + "All Reads\tALL\t20\t3735\t37\t14\t3.698163\t0.607974\t\t\t", + "All Reads\tALL\t21\t3759\t40\t14\t3.972488\t0.628105\t\t\t", + "All Reads\tALL\t22\t3461\t31\t14\t3.343759\t0.600557\t\t\t", + "All Reads\tALL\t23\t3475\t31\t13\t3.330288\t0.598137\t\t\t", + "All Reads\tALL\t24\t3393\t27\t13\t2.970673\t0.571706\t\t\t", + "All Reads\tALL\t25\t3365\t44\t12\t4.881379\t0.735896\t\t\t", + "All Reads\tALL\t26\t3200\t27\t12\t3.149841\t0.606187\t\t\t", + "All Reads\tALL\t27\t2988\t33\t13\t4.122952\t0.717714\t\t\t", + "All Reads\tALL\t28\t2852\t37\t12\t4.843141\t0.796207\t\t\t", + "All Reads\tALL\t29\t2805\t38\t13\t5.057381\t0.820415\t\t\t", + "All Reads\tALL\t30\t2833\t21\t12\t2.767245\t0.603862\t\t\t", + "All Reads\tALL\t31\t2731\t19\t10\t2.597208\t0.59584\t\t\t", + "All Reads\tALL\t32\t2760\t18\t11\t2.43466\t0.573855\t\t\t", + "All Reads\tALL\t33\t2776\t23\t13\t3.093024\t0.64494\t\t\t", + "All Reads\tALL\t34\t2475\t12\t14\t1.81001\t0.522505\t\t\t", + "All Reads\tALL\t35\t2672\t8\t13\t1.117708\t0.39517\t\t\t", + "All Reads\tALL\t36\t2796\t6\t12\t0.801104\t0.327049\t\t\t", + "All Reads\tALL\t37\t2850\t13\t11\t1.702838\t0.472282\t\t\t", + "All Reads\tALL\t38\t2826\t5\t12\t0.6605\t0.295385\t\t\t", + "All Reads\tALL\t39\t3054\t4\t13\t0.488952\t0.244476\t\t\t", + "All Reads\tALL\t40\t3449\t0\t0\t0\t0\t\t\t", + "All Reads\tALL\t41\t4012\t6\t13\t0.558297\t0.227924\t\t\t", + "All Reads\tALL\t42\t4971\t5\t12\t0.375492\t0.167925\t\t\t", + "All Reads\tALL\t43\t5630\t1\t13\t0.066308\t0.066308\t\t\t", + "All Reads\tALL\t44\t6409\t0\t0\t0\t0\t\t\t", + "All Reads\tALL\t45\t7297\t1\t12\t0.05116\t0.05116\t\t\t", + "All Reads\tALL\t46\t8146\t1\t8\t0.045828\t0.045828\t\t\t", + "All Reads\tALL\t47\t8876\t2\t8\t0.084118\t0.05948\t\t\t", "All Reads\tALL\t48\t9213\t0\t0\t0\t0\t\t\t", "All Reads\tALL\t49\t9815\t0\t0\t0\t0\t\t\t", - "All Reads\tALL\t50\t10134\t1\t6\t0.025529\t0.025529\t\t\t", - "All Reads\tALL\t51\t9913\t1\t3\t0.026098\t0.026098\t\t\t", - "All Reads\tALL\t52\t9189\t1\t2\t0.028154\t0.028154\t\t\t", + "All Reads\tALL\t50\t10134\t1\t22\t0.036838\t0.036838\t\t\t", + "All Reads\tALL\t51\t9913\t0\t0\t0\t0\t\t\t", + "All Reads\tALL\t52\t9189\t0\t0\t0\t0\t\t\t", "All Reads\tALL\t53\t8455\t0\t0\t0\t0\t\t\t", "All Reads\tALL\t54\t7312\t0\t0\t0\t0\t\t\t", "All Reads\tALL\t55\t5984\t0\t0\t0\t0\t\t\t", @@ -230,18 +218,18 @@ "alignment_metrics", [ "CATEGORY\tTOTAL_READS\tPF_READS\tPCT_PF_READS\tPF_NOISE_READS\tPF_READS_ALIGNED\tPCT_PF_READS_ALIGNED\tPF_ALIGNED_BASES\tPF_HQ_ALIGNED_READS\tPF_HQ_ALIGNED_BASES\tPF_HQ_ALIGNED_Q20_BASES\tPF_HQ_MEDIAN_MISMATCHES\tPF_MISMATCH_RATE\tPF_HQ_ERROR_RATE\tPF_INDEL_RATE\tMEAN_READ_LENGTH\tSD_READ_LENGTH\tMEDIAN_READ_LENGTH\tMAD_READ_LENGTH\tMIN_READ_LENGTH\tMAX_READ_LENGTH\tMEAN_ALIGNED_READ_LENGTH\tREADS_ALIGNED_IN_PAIRS\tPCT_READS_ALIGNED_IN_PAIRS\tPF_READS_IMPROPER_PAIRS\tPCT_PF_READS_IMPROPER_PAIRS\tBAD_CYCLES\tSTRAND_BALANCE\tPCT_CHIMERAS\tPCT_ADAPTER\tPCT_SOFTCLIP\tPCT_HARDCLIP\tAVG_POS_3PRIME_SOFTCLIP_LENGTH\tSAMPLE\tLIBRARY\tREAD_GROUP", - "UNPAIRED\t1100\t1094\t0.994545\t0\t1094\t1\t80799\t224\t16598\t15805\t0\t0.205196\t0.161755\t0.004616\t75.754113\t2.037807\t76\t0\t44\t76\t74.196527\t0\t0\t0\t0\t0\t0.674589\t0.004464\t0\t0.020561\t0\t18.139535\t\t\t", + "UNPAIRED\t745\t745\t1\t0\t745\t1\t99162\t537\t76242\t75002\t0\t0.034304\t0.014897\t0.002219\t148.559732\t6.956237\t150\t0\t71\t150\t133.465772\t0\t0\t0\t0\t0\t0.989262\t0.018622\t0\t0.101602\t0\t49.564103\t\t\t", "## HISTOGRAM\tjava.lang.Integer", "READ_LENGTH\tUNPAIRED_TOTAL_LENGTH_COUNT\tUNPAIRED_ALIGNED_LENGTH_COUNT" ] ], [ "methyldackel_extract", - "91eac7636f2e617927f190e3e884b8c9" + "55ab0743f01e4d66921bacd7dd3ef411" ], [ "mbias", - "824e46917dfa852f1f3ed83571008eef" + "20c46a85d89d9fa53ed671a1372a7299" ], [ "nonconverted", @@ -252,15 +240,17 @@ [ "nonconverted_combined", [ - "emseq-test1\tchr1\t0" + "emseq-test1.ds.1\tchr1\t0" ] ], [ "positional_summary", [ "methylkit_file\tchr\ttarget_length\tposition\tcontext\tmean_methylation\tn_loci\tn_measurements", - "emseq-test1\tchr1\t182\t178\tCG\t0\t1\t1", - "emseq-test1\tchr1\t204\t189\tCG\t0\t1\t1" + "emseq-test1.ds.1\tchr1\t208\t110\tCG\t100\t1\t1", + "emseq-test1.ds.1\tchr1\t204\t189\tCG\t33.3333\t1\t1", + "emseq-test1.ds.1\tchr1\t151\t142\tCG\t100\t1\t1", + "emseq-test1.ds.1\tchr1\t182\t178\tCG\t33.3333\t1\t1" ] ], [ @@ -270,17 +260,13 @@ [ "region_summary", [ - "library\ttarget_locus\ttarget_name\tcontext\tmean_methylation", - "emseq-test2\tchr1:140839-141043\tStemLoop_140889_140992_2mm_22C_0CG\tCG\t0", - "emseq-test2\tchr1:140850-141032\tStemLoop_140900_140981_0mm_29C_1CG\tCG\t0" + "library\ttarget_locus\ttarget_name\tcontext\tmean_methylation" ] ], [ "methylkit_intersections", [ - "methylkit_file\tchr\tstart\tend\tcontext\tmethylation\ttarget_locus\ttarget_name", - "emseq-test2\tchr1\t141027\t141028\tCG\t0\tchr1:140840-141043\tStemLoop_140889_140992_2mm_22C_0CG", - "emseq-test2\tchr1\t141027\t141028\tCG\t0\tchr1:140851-141032\tStemLoop_140900_140981_0mm_29C_1CG" + "methylkit_file\tchr\tstart\tend\tcontext\tmethylation\ttarget_locus\ttarget_name" ] ], [ @@ -294,36 +280,36 @@ [ "ngs_agg", [ - "--bam emseq-test1.md.bam ", - "--bai emseq-test1.md.bai ", - "--metadata_bam_file emseq-test1.bam ", - "--fastp emseq-test1.fastp.json ", - "--aln emseq-test1.alignment_summary_metrics.txt ", - "--gc emseq-test1.gc_metrics ", - "--dup emseq-test1.markdups_log ", - "--idx_stats emseq-test1.idxstat ", - "--flagstat emseq-test1.flagstat ", - "--fastqc emseq-test1.md_fastqc.zip ", - "--nonconverted_read_counts emseq-test1.nonconverted_counts.for_agg.tsv ", - "--tasmanian emseq-test1.tasmanian.csv ", - "--combined_mbias_records emseq-test1.combined_mbias.tsv ", - "--fgbio_switchbacks emseq-test1.switchbacks.summary.txt", + "--bam emseq-test1.ds.1.md.bam ", + "--bai emseq-test1.ds.1.md.bai ", + "--metadata_bam_file emseq-test1.ds.1.bam ", + "--fastp emseq-test1.ds.1.fastp.json ", + "--aln emseq-test1.ds.1.alignment_summary_metrics.txt ", + "--gc emseq-test1.ds.1.gc_metrics ", + "--dup emseq-test1.ds.1.markdups_log ", + "--idx_stats emseq-test1.ds.1.idxstat ", + "--flagstat emseq-test1.ds.1.flagstat ", + "--fastqc emseq-test1.ds.1.md_fastqc.zip ", + "--nonconverted_read_counts emseq-test1.ds.1.nonconverted_counts.for_agg.tsv ", + "--tasmanian emseq-test1.ds.1.tasmanian.csv ", + "--combined_mbias_records emseq-test1.ds.1.combined_mbias.tsv ", + "--fgbio_switchbacks emseq-test1.ds.1.switchbacks.summary.txt", "--workflow EM-seq" ] ] ], + "timestamp": "2026-09-06T00:59:18.912472896", "meta": { - "nf-test": "0.9.5", - "nextflow": "24.10.5" - }, - "timestamp": "2026-08-20T15:14:15.067200906" + "nf-test": "0.9.4", + "nextflow": "25.10.4" + } }, "EM-seq workflow uBam": { "content": [ { - "tasksCount": 49, + "tasksCount": 45, "tasksFailed": 0, - "tasksSucceeded": 49 + "tasksSucceeded": 45 }, [ "bwameth_file_1", @@ -357,7 +343,7 @@ "fastp", [ "\t\"summary\": {", - "\t\t\"fastp_version\": \"1.3.6\",", + "\t\t\"fastp_version\": \"0.1.0\",", "\t\t\"sequencing\": \"paired end (76 cycles + 76 cycles)\",", "\t\t\"before_filtering\": {", "\t\t\t\"total_reads\": 9534,", @@ -368,18 +354,18 @@ "\t\t\t\"q30_rate\": 0.913898,", "\t\t\t\"read1_mean_length\": 76,", "\t\t\t\"read2_mean_length\": 76,", - "\t\t\t\"gc_content\": 0.218667", + "\t\t\t\"gc_content\": 0.218668", "\t\t},", "\t\t\"after_filtering\": {", "\t\t\t\"total_reads\": 9526,", - "\t\t\t\"total_bases\": 719151,", - "\t\t\t\"q20_bases\": 676526,", - "\t\t\t\"q30_bases\": 657902,", - "\t\t\t\"q20_rate\": 0.940729,", - "\t\t\t\"q30_rate\": 0.914832,", + "\t\t\t\"total_bases\": 719317,", + "\t\t\t\"q20_bases\": 676675,", + "\t\t\t\"q30_bases\": 658048,", + "\t\t\t\"q20_rate\": 0.940719,", + "\t\t\t\"q30_rate\": 0.914823,", "\t\t\t\"read1_mean_length\": 75,", "\t\t\t\"read2_mean_length\": 75,", - "\t\t\t\"gc_content\": 0.216787", + "\t\t\t\"gc_content\": 0.216795", "\t\t}", "\t},", "\t\"filtering_result\": {", @@ -429,20 +415,20 @@ [ "flagstats", [ - "1215 + 584 in total (QC-passed reads + QC-failed reads)", - "1214 + 584 primary", + "1799 + 0 in total (QC-passed reads + QC-failed reads)", + "1798 + 0 primary", "1 + 0 secondary", "0 + 0 supplementary", - "22 + 2 duplicates", - "22 + 2 primary duplicates", - "956 + 579 mapped (78.68% : 99.14%)", - "955 + 579 primary mapped (78.67% : 99.14%)", - "1214 + 584 paired in sequencing", - "607 + 292 read1", - "607 + 292 read2", - "674 + 0 properly paired (55.52% : 0.00%)", - "696 + 574 with itself and mate mapped", - "259 + 5 singletons (21.33% : 0.86%)", + "24 + 0 duplicates", + "24 + 0 primary duplicates", + "1535 + 0 mapped (85.33% : N/A)", + "1534 + 0 primary mapped (85.32% : N/A)", + "1798 + 0 paired in sequencing", + "899 + 0 read1", + "899 + 0 read2", + "674 + 0 properly paired (37.49% : N/A)", + "1270 + 0 with itself and mate mapped", + "264 + 0 singletons (14.68% : N/A)", "0 + 0 with mate mapped to a different chr", "0 + 0 with mate mapped to a different chr (mapQ>=5)" ] @@ -558,9 +544,9 @@ "alignment_metrics", [ "CATEGORY\tTOTAL_READS\tPF_READS\tPCT_PF_READS\tPF_NOISE_READS\tPF_READS_ALIGNED\tPCT_PF_READS_ALIGNED\tPF_ALIGNED_BASES\tPF_HQ_ALIGNED_READS\tPF_HQ_ALIGNED_BASES\tPF_HQ_ALIGNED_Q20_BASES\tPF_HQ_MEDIAN_MISMATCHES\tPF_MISMATCH_RATE\tPF_HQ_ERROR_RATE\tPF_INDEL_RATE\tMEAN_READ_LENGTH\tSD_READ_LENGTH\tMEDIAN_READ_LENGTH\tMAD_READ_LENGTH\tMIN_READ_LENGTH\tMAX_READ_LENGTH\tMEAN_ALIGNED_READ_LENGTH\tREADS_ALIGNED_IN_PAIRS\tPCT_READS_ALIGNED_IN_PAIRS\tPF_READS_IMPROPER_PAIRS\tPCT_PF_READS_IMPROPER_PAIRS\tBAD_CYCLES\tSTRAND_BALANCE\tPCT_CHIMERAS\tPCT_ADAPTER\tPCT_SOFTCLIP\tPCT_HARDCLIP\tAVG_POS_3PRIME_SOFTCLIP_LENGTH\tSAMPLE\tLIBRARY\tREAD_GROUP", - "FIRST_OF_PAIR\t899\t607\t0.675195\t0\t493\t0.812191\t32309\t302\t19684\t18772\t3\t0.103312\t0.098954\t0.004364\t74.663921\t5.91042\t76\t0\t31\t76\t53.443163\t348\t0.705882\t156\t0.31643\t0\t0.89858\t0.010101\t0\t0.093312\t0\t15.835294\t\t\t", - "SECOND_OF_PAIR\t899\t607\t0.675195\t0\t462\t0.76112\t30277\t299\t19999\t18469\t3\t0.102474\t0.09557\t0.004096\t74.710049\t5.864669\t76\t0\t31\t76\t50.116969\t348\t0.753247\t125\t0.270563\t0\t0.112554\t0.010453\t0\t0.086176\t0\t14.56701\t\t\t", - "PAIR\t1798\t1214\t0.675195\t0\t955\t0.786656\t62586\t601\t39683\t37241\t3\t0.102909\t0.097258\t0.004234\t74.686985\t5.885207\t76\t0\t31\t76\t51.780066\t696\t0.728796\t281\t0.294241\t0\t0.518325\t0.010274\t0\t0.089743\t0\t15.159341\t\t\t" + "FIRST_OF_PAIR\t899\t899\t1\t0\t781\t0.868743\t42522\t344\t22297\t21272\t0\t0.109413\t0.102292\t0.005362\t74.83426\t5.220772\t76\t0\t31\t76\t47.575083\t635\t0.81306\t444\t0.568502\t0\t0.806658\t0.010101\t0\t0.231137\t0\t23.036866\t\t\t", + "SECOND_OF_PAIR\t899\t899\t1\t3\t753\t0.837597\t40929\t343\t22862\t21179\t0\t0.110193\t0.097406\t0.004984\t74.865406\t5.184858\t76\t0\t31\t76\t45.799778\t635\t0.843293\t416\t0.552457\t0\t0.203187\t0.010453\t0\t0.223375\t0\t20.209016\t\t\t", + "PAIR\t1798\t1798\t1\t3\t1534\t0.85317\t83451\t687\t45159\t42451\t0\t0.109795\t0.09983\t0.005177\t74.849833\t5.201422\t76\t0\t31\t76\t46.68743\t1270\t0.827901\t860\t0.560626\t0\t0.51043\t0.010274\t0\t0.227255\t0\t21.54013\t\t\t" ] ], [ @@ -674,10 +660,10 @@ ] ] ], + "timestamp": "2026-09-06T14:50:43.584038567", "meta": { - "nf-test": "0.9.5", - "nextflow": "24.10.5" - }, - "timestamp": "2026-08-20T15:13:32.113269349" + "nf-test": "0.9.4", + "nextflow": "25.10.4" + } } } \ No newline at end of file