In-depth mutation and SNP discovery using DHPLC gene scanning

Abstract

Denaturing high-performance liquid chromatography (DHPLC) is a new technology used in the discovery of genetic variations (mutations), such as single-base substitutions (or single nucleotide polymorphisms) and small deletions or insertions. These genetic variations can be routinely detected by DHPLC gene scanning at the germ-line and somatic levels. Epigenetic alterations, such as changes in DNA methylation status at defined loci, can also be assessed using DHPLC-based methodologies. The biological impact of these genetic variations depends on the location and identity of the DNA sequence alteration. The discovery of functionally relevant genetic variations can be exploited throughout the drug discovery and development processes. Examples of the application of DHPLC for sequence variant detection will be presented and discussed, with emphasis on target validation by candidate gene scanning and mutation detection in disease pathway genes, as well as the discovery of therapeutically significant mutations associated with drug metabolism and resistance.