Bio Vcf Basics

VCF/BCF Basics

View and query variant files using bcftools and cyvcf2.

Format Overview

VCF Format Structure

##fileformat=VCFv4.2
##INFO=<ID=DP,Number=1,Type=Integer,Description="Total Depth">
##FORMAT=<ID=GT,Number=1,Type=String,Description="Genotype">
##FORMAT=<ID=DP,Number=1,Type=Integer,Description="Read Depth">
#CHROM  POS     ID      REF     ALT     QUAL    FILTER  INFO    FORMAT  SAMPLE1
chr1    1000    rs123   A       G       30      PASS    DP=50   GT:DP   0/1:25

Header Lines (##)

• ##fileformat - VCF version
• ##INFO - INFO field definitions
• ##FORMAT - FORMAT field definitions
• ##FILTER - Filter definitions
• ##contig - Reference contigs
• ##reference - Reference genome

Column Header (#CHROM)

Fixed columns: CHROM, POS, ID, REF, ALT, QUAL, FILTER, INFO, FORMAT Followed by sample columns

Data Columns

bcftools view

View VCF

bcftools view input.vcf.gz | head

View Header Only

bcftools view -h input.vcf.gz

View Without Header

bcftools view -H input.vcf.gz | head

View Specific Region

bcftools view input.vcf.gz chr1:1000000-2000000

View Specific Samples

bcftools view -s sample1,sample2 input.vcf.gz

Exclude Samples

bcftools view -s ^sample3 input.vcf.gz

bcftools query

Extract specific fields in custom format.

Basic Query

bcftools query -f '%CHROM\t%POS\t%REF\t%ALT\n' input.vcf.gz

Query with INFO Fields

bcftools query -f '%CHROM\t%POS\t%INFO/DP\t%INFO/AF\n' input.vcf.gz

Query with Sample Fields

bcftools query -f '%CHROM\t%POS[\t%GT]\n' input.vcf.gz

Query Specific Samples

bcftools query -f '%CHROM\t%POS[\t%SAMPLE=%GT]\n' -s sample1,sample2 input.vcf.gz

Include Header

bcftools query -H -f '%CHROM\t%POS\t%REF\t%ALT\n' input.vcf.gz

Common Format Specifiers

Format Conversion

VCF to BCF

bcftools view -Ob -o output.bcf input.vcf.gz

BCF to VCF

bcftools view -Ov -o output.vcf input.bcf

Compress VCF (bgzip)

bgzip input.vcf
# Creates input.vcf.gz

Index VCF/BCF

bcftools index input.vcf.gz
# Creates input.vcf.gz.csi

bcftools index -t input.vcf.gz
# Creates input.vcf.gz.tbi (tabix index)

Output Format Options

Genotype Encoding

cyvcf2 Python Alternative

Open and Iterate

from cyvcf2 import VCF

vcf = VCF('input.vcf.gz')
for variant in vcf:
    print(f'{variant.CHROM}:{variant.POS} {variant.REF}>{variant.ALT[0]}')

Access Variant Properties

from cyvcf2 import VCF

for variant in VCF('input.vcf.gz'):
    print(f'Chrom: {variant.CHROM}')
    print(f'Pos: {variant.POS}')
    print(f'ID: {variant.ID}')
    print(f'Ref: {variant.REF}')
    print(f'Alt: {variant.ALT}')  # List
    print(f'Qual: {variant.QUAL}')
    print(f'Filter: {variant.FILTER}')
    print(f'Type: {variant.var_type}')  # snp, indel, etc.
    break

Access INFO Fields

from cyvcf2 import VCF

for variant in VCF('input.vcf.gz'):
    dp = variant.INFO.get('DP')
    af = variant.INFO.get('AF')
    print(f'{variant.CHROM}:{variant.POS} DP={dp} AF={af}')

Access Genotypes

from cyvcf2 import VCF

vcf = VCF('input.vcf.gz')
samples = vcf.samples  # List of sample names

for variant in vcf:
    gts = variant.gt_types  # 0=HOM_REF, 1=HET, 2=UNKNOWN, 3=HOM_ALT
    for sample, gt in zip(samples, gts):
        gt_str = ['HOM_REF', 'HET', 'UNKNOWN', 'HOM_ALT'][gt]
        print(f'{sample}: {gt_str}')
    break

Access Sample Fields

from cyvcf2 import VCF

for variant in VCF('input.vcf.gz'):
    depths = variant.format('DP')  # numpy array
    gqs = variant.format('GQ')     # Genotype quality
    print(f'Depths: {depths}')

Fetch Region

from cyvcf2 import VCF

vcf = VCF('input.vcf.gz')
for variant in vcf('chr1:1000000-2000000'):
    print(f'{variant.CHROM}:{variant.POS}')

Get Header Info

from cyvcf2 import VCF

vcf = VCF('input.vcf.gz')
print(f'Samples: {vcf.samples}')
print(f'Contigs: {vcf.seqnames}')

# INFO field definitions
for info in vcf.header_iter():
    if info['HeaderType'] == 'INFO':
        print(f'{info["ID"]}: {info["Description"]}')

Write VCF

from cyvcf2 import VCF, Writer

vcf = VCF('input.vcf.gz')
writer = Writer('output.vcf', vcf)

for variant in vcf:
    if variant.QUAL > 30:
        writer.write_record(variant)

writer.close()
vcf.close()

Quick Reference

Common Errors

Related Skills

• variant-calling - Generate VCF from alignments
• filtering-best-practices - Filter variants by quality/criteria
• vcf-manipulation - Merge, concat, intersect VCFs
• alignment-files/pileup-generation - Generate pileup for calling