Abstract
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.
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Acknowledgements
DJM and VMH are Cancer Institute NSW Fellows (Australia). The Humpty Dumpty Foundation, Australia, is acknowledged for the donation of DHPLC equipment.
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Marsh, D.J., Howell, V.M. (2010). The Use of Denaturing High Performance Liquid Chromatography (DHPLC) for Mutation Scanning of Hereditary Cancer Genes. In: Webb, M. (eds) Cancer Susceptibility. Methods in Molecular Biology, vol 653. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-759-4_8
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DOI: https://doi.org/10.1007/978-1-60761-759-4_8
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