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Samplesheet Format

The Nextflow pipeline accepts a CSV samplesheet with the following columns:

Columns

sample,bam,mfsd_bam,meth_bam,vcf,bed,maf,single_sample_maf,assay,pon,targets
Column Type Required Description
sample TEXT Sample identifier
bam PATH WGS/Panel BAM file (all_unique for panels)
mfsd_bam PATH Duplex BAM for mFSD (falls back to bam if empty)
meth_bam PATH Bisulfite BAM for UXM
vcf PATH VCF for mFSD
bed PATH Pre-extracted .bed.gz
maf PATH MAF for mFSD
single_sample_maf BOOL Skip MAF filtering if true
assay TEXT Assay code: XS1, XS2, WGS
pon PATH Sample-specific PON (overrides assay)
targets PATH Sample-specific targets (overrides assay)

Input Logic

Input Combination Workflow
bam only Full run-all (extract → features)
bam + vcf/maf run-all + mFSD
bam + mfsd_bam + maf run-all with dual BAM support
meth_bam only UXM methylation
bed only FSC, FSR, FSD, WPS, OCF (no extract)

Assay Resolution

When assay is set and --asset_dir is provided:

Assay PON Targets Genes
XS1 xs1.pon.parquet xs1.targets.bed 128
XS2 xs2.pon.parquet xs2.targets.bed 146
WGS wgs.pon.parquet None -

Example

sample,bam,mfsd_bam,meth_bam,vcf,bed,maf,single_sample_maf,assay,pon,targets
# MSK-ACCESS V1 samples (auto-resolve PON/targets)
ACCESS_001,/data/sample1.bam,,,,,,,XS1,,
ACCESS_002,/data/sample2.bam,,,,,,/data/cohort.maf,false,XS2,,

# MSK-ACCESS V2 with duplex BAM for mFSD
ACCESS_V2,/data/sample.all_unique.bam,/data/sample.duplex.bam,,,,,maf.tsv,true,XS2,,

# WGS (no targets)
WGS_001,/data/wgs.bam,,,/data/wgs.vcf,,,,,WGS,,

# Custom PON/targets
CUSTOM,/data/custom.bam,,,,,,,,,/data/custom.pon.parquet,/data/custom.bed