Description and validation of high-throughput simultaneous genotyping and mutation scanning by high-resolution melting curve analysis

摘要: Mutation scanning using high-resolution melt- ing curve analysis (HR-melt) is an effective and sensitive method to detect sequence variations. However, the presence of a common SNP within mutation amplicon may considerably complicate interpretation results increase number samples flagged for sequencing by interfering with clustering according melting profiles. A protocol describing simultaneous gene genotyp- has been reported. Here, we show that it can improve sensitivity efficiency large-scale case-control screening. Two exons ATM, both containing standard scanning, were selected screening 1,356 subjects from interna- tional breast cancer genetics study. Asymmetric PCR was performed in SNP-specific unlabeled probe. Stratification their probe-target aided customized HR-melt software. This approach improved identification rare known unknown variants, while dramatically reducing effort. It even allowed genotyping tandem SNPs single Hence, rapid, efficient, cost-effective tool be used high-throughput research, as well molecular diagnostic clinical purposes. Hum Mutat 30, 884-890, 2009. & 2009 Wiley-Liss, Inc.