Fill Ref Md5

fill-ref-md5

Quick Start

• Command: fill-ref-md5 [OPTIONS] in.vcf.gz out.vcf.gz
• Local executable: /home/vimalinx/miniforge3/envs/bio/bin/fill-ref-md5
• Reference: see references/help.md for full options and examples

When To Use This Tool

• Add reference and contig header lines with MD5 checksums to a compressed VCF.
• Make a VCF header more compliant with the VCFv4.1-style reference metadata recommendations.
• Populate missing contig MD5 values from a reference FASTA before data sharing or downstream indexing.

Common Patterns

# 1) Compute MD5s from a reference FASTA and cache them in a dictionary
fill-ref-md5 -r ref.fa -d ref.fa.dict in.vcf.gz out.vcf.gz

# 2) Add assembly/species/taxonomy annotations to contig header lines
fill-ref-md5 \
  -r ref.fa \
  -d ref.fa.dict \
  -i AS:GRCh38,SP:"Homo sapiens",TX:9606 \
  in.vcf.gz out.vcf.gz

Recommended Workflow

1. Ensure input VCF is bgzip-compressed and tabix-indexed
2. Prepare reference FASTA indexed by samtools faidx
3. Run fill-ref-md5 -r ref.fa -d ref.fa.dict in.vcf.gz out.vcf.gz
4. Verify output VCF header contains new reference/contig tags

Guardrails

• Input VCF must be compressed and tabix-indexed
• Reference FASTA must be indexed by samtools faidx
• Dictionary file opens in append mode; existing records are not modified
• You need at least one of -d or -r; -d alone only works if the dictionary already contains all required chromosomes.
• --help works, but --version is not implemented and errors as an unknown parameter.
• The script shells out to tabix, samtools faidx, and md5sum, so those helpers must be available.