Journal article
A high-throughput protocol for mutation scanning of the BRCA1 and BRCA2 genes
HL Hondow, SB Fox, G Mitchell, RJ Scott, V Beshay, SQ Wong, A Dobrovic
BMC Cancer | BIOMED CENTRAL LTD | Published : 2011
Open access
Abstract
Background: Detection of mutations by DNA sequencing can be facilitated by scanning methods to identify amplicons which may have mutations. Current scanning methods used for the detection of germline sequence variants are laborious as they require post-PCR manipulation. High resolution melting (HRM) is a cost-effective rapid screening strategy, which readily detects heterozygous variants by melting curve analysis of PCR products. It is well suited to screening genes such as BRCA1 and BRCA2 as germline pathogenic mutations in these genes are always heterozygous.Methods: Assays for the analysis of all coding regions and intron-exon boundaries of BRCA1 and BRCA2 were designed, and optimised. A ..
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Funding Acknowledgements
This project was supported by a Priority Driven Collaborative Cancer Research Scheme grant from Cancer Australia partnered by the National Breast Cancer Foundation and the Prostate Cancer Foundation of Australia to AD, SF and GM and a New Technology grant from the Victorian Department of Human Services to SF, GM and AD. We thank Trent Warburton from Trendbio Sciences for technical discussion. We especially thank our consumer representative, Gerda Evans for her support. We thank Hongdo Do for his critical reading of the final version of the manuscript. We thank Cliff Meldrum (Peter MacCallum Cancer Centre) and Margie Smith and Emanouil Sigalas (Royal Melbourne Hospital) for DNA samples used in this study. We also thank Amber Willems and Heather Thorne from kConFab for positive control DNA samples. We thank the kConFab research nurses and staff, the heads and staff of the Family Cancer Clinics, and the Clinical Follow Up Study for their contributions to the kConFab resource, and the many families who contribute to kConFab. kConFab is supported by grants from the National Breast Cancer Foundation, the National Health and Medical Research Council of Australia, the Queensland Cancer Fund, the Cancer Councils of New South Wales, Victoria, Tasmania and South Australia, and the Cancer Foundation of Western Australia.