The Use of Denaturing High Performance Liquid Chromatography (DHPLC) for Mutation Scanning of Heredi

Denaturing high performance liquid chromatography (DHPLC) facilitates automated mutation scanning of PCR products with the ability to detect nearly 100% of sequence variants including single nucleotide substitutions and small insertions or deletions. It has particular application for genetic screening in inherited conditions; both for the initial identification of a mutation in disease carriers followed by sequence analysis, and for screening “at-risk” individuals prior to the development of disease in families with a known mutation. Specifically, in familial cancer syndromes, DHPLC has been reported as a genetic screening tool for the risk of developing breast and ovarian cancer (BRCA1 ), von Hippel Lindau disease (VHL ), Cowden syndrome (PTEN ), and Multiple Endocrine Neoplasia types 1 and 2 (MEN1 and RET ). This chapter focuses on the methodologies specific to the WAVE� System for Mutation Detection 2100 (Transgenomic Inc., Omaha, NE, USA) and highlights the use of Navigator™ software (Transgenomic Inc.), including data analysis with scatter graphs.