Use of DNA thermodynamics for low-abundance mutation detection by DNA hybridization

Abstract

Duplex formation via hybridization provides a detection technique where one of the strands is designed to match a mutant sequence and probe its presence. In a majority wild-type background consisting of single-nucleotide variations, the probe is however subject to cross-hybridization, lowering the detection sensitivity of low-abundant mutants. We use thermodynamics-based probe design and Langmuir theory to increase sensitivity and quantify the amount of mutant in a sample.