A high-throughput protocol for mutation scanning of the BRCA1 and BRCA2 genes.

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 final set of 94 assays which ran under identical amplification conditions were chosen

SUBMITTER: Hondow HL 

PROVIDER: S-EPMC3146935 | biostudies-literature | 2011 Jun

REPOSITORIES: biostudies-literature