Mutation Scanning for the Clinical Laboratory: DHPLC

This chapter describes the Transgenomic WAVE� DNA Fragment Analysis System with DNA Sep� Technology, as used in our molecular genetics diagnostic laboratories for the detection of unknown mutations. Four software packages are currently available: WAVEmaker 3.4, 4, 4.1, and most recently, Navigator. Because versions 4 and 4.1 are similar, version 4 is presented here. Denaturing high-performance liquid chromatography (DHPLC), also known as TMHA (temperature-modulated heteroduplex analysis) provides a rapid, automated scanning method for mutations, whether known or unknown. Mutations are visualized as a characteristic pattern of peaks corresponding to the mixture of heteroduplexes and homoduplexes formed when wild-type and mutant DNA fragments are hybridized (1 ; Fig. 1 ).

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