Rare Variant Quantitation Using Droplet Digital PCR

Abstract

Droplet digital PCR (ddPCR) allows precise quantitation of individual copies of DNA. Using allele-specific fluorescent oligonucleotide probes, ddPCR can enumerate both mutant and wild-type alleles enabling highly sensitive detection and quantitation of the variant allele frequency of mutated genes. In this protocol, we describe a method for using ddPCR to detect mutations in genomic DNA with a sensitivity of up to 1 in 50,000 DNA copies.