Allele Balance in VCF Calculator

Measure allele balance from VCF fields. Compare ref and alternate support. Use custom balance thresholds. Flag biased calls, low depth, and genotype evidence quickly.

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Calculator

Example Data Table

Case GT AD DP ALT Balance Meaning
Balanced heterozygous 0/1 18,12 30 0.4000 Inside a common heterozygous review range.
Low alternate support 0/1 29,3 32 0.0938 Needs review for weak alternate evidence.
High alternate support 1/1 2,40 42 0.9524 Fits strong alternate read support.

Formula Used

Selected allele balance = ALT reads / (REF reads + selected ALT reads).

Reference balance = REF reads / (REF reads + selected ALT reads).

Variant allele fraction = selected ALT reads / total depth DP.

All alternate balance = sum of all ALT reads / sum of AD reads.

Distance from expected = absolute value of selected allele balance minus expected balance.

How to Use This Calculator

Enter reference reads and alternate reads. Add total depth if available. You can also paste an AD string, such as 18,12. For a full sample column, paste FORMAT and SAMPLE values together. Choose the alternate index when the record has several alternate alleles. Set depth and balance limits. Press Submit. The result appears above the form.

Why Allele Balance Matters

Allele balance shows how much read support belongs to an alternate allele. It is often checked in VCF review. A balanced heterozygous call usually has similar reference and alternate read support. A very low balance can suggest noise, mapping bias, strand issues, sample mixture, or a weak call. The calculator helps you inspect those values before deeper review.

Reading VCF Depth Fields

VCF files often store read counts in the AD field. The first AD value usually represents reference reads. Later values represent alternate alleles. For a biallelic site, AD=18,12 means 18 reference reads and 12 alternate reads. The alternate allele balance is 12 divided by 30. That gives 0.4000, or 40.00 percent.

Flexible Input Options

This tool also accepts FORMAT and SAMPLE strings. It can read AD, DP, GT, and GQ when those keys exist. This is useful when copying one sample column from a VCF line. You may also enter read counts manually. Manual fields are helpful when the source uses a custom tag.

Review Thresholds

Use the minimum depth filter to catch thin evidence. Use the balance range to set your review limits. A common heterozygous review range is 0.30 to 0.70. That range is not universal. Tumor samples, pooled samples, mosaic sites, copy number changes, and contamination can shift expected balance. Always compare results with your assay design and quality rules.

Understanding the Report

The report shows alternate balance, reference balance, variant allele fraction, total depth, selected allele support, and status. It also explains which input source was used. This reduces confusion when several VCF fields are present.

Advanced Review Notes

Advanced review needs context. A homozygous alternate genotype should trend toward high alternate support. A reference genotype should trend toward low alternate support. Multi allelic records need the correct alternate index. Low depth can make any ratio unstable. High depth can still be biased by alignment or chemistry effects.

Exporting Results

Exports are useful for batch notes. CSV files work well in spreadsheets. PDF reports are useful for summaries, reviews, and internal records. The result should not replace expert variant interpretation. It is a quick numeric check for read support. Confirm flagged variants with the full VCF, alignment view, filters, and laboratory standards. Save the chosen thresholds with each export. This makes repeat review easier later too.

FAQs

What is allele balance in a VCF file?

Allele balance is the fraction of read support for an allele. For a selected alternate allele, it is usually ALT reads divided by REF plus ALT reads.

Which VCF field is used for read counts?

The AD field is commonly used. It usually lists reference read count first, followed by alternate allele read counts. Some callers may use other tags.

How do I enter a multi allelic AD value?

Paste the full AD list, such as 22,10,4. Then set the alternate allele index. Index 1 means the first alternate allele after reference.

What range is normal for heterozygous calls?

A common review range is 0.30 to 0.70. It is not a universal rule. Assay type, sample purity, and variant context matter.

What does low alternate balance mean?

It means alternate support is weak compared with reference support. This may indicate noise, mapping bias, low fraction variants, or an uncertain call.

Can I paste FORMAT and SAMPLE values?

Yes. Enter FORMAT like GT:AD:DP:GQ. Then enter the matching SAMPLE value. The calculator reads supported keys from those fields.

How is VAF different from allele balance?

VAF uses selected alternate reads divided by total depth. Allele balance uses selected alternate reads divided by reference plus selected alternate reads.

Should this replace variant interpretation?

No. It is a numeric screening aid. Review the full VCF, alignments, caller filters, sample context, and laboratory rules before making decisions.

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Important Note: All the Calculators listed in this site are for educational purpose only and we do not guarentee the accuracy of results. Please do consult with other sources as well.