3250 LC 480 TN 1.indd

High resolution melting (HRM) is a novel, closed-tube, post-PCR technique allowing genomic researchers to easily analyze genetic variations in PCR amplicons. This technical note describes general steps of setting up HRM-based PCR assays, with a special focus on ways how to optimize procedures for gene scanning experiments. The novel High Resolution Melting (HRM) technique enables researchers to rapidly and efficiently discover genetic variations (e.g., SNPs, mutations, methylations). HRM provides outstanding specific-ity and sensitivity with high sample throughput. Using the LightCycler® 480 Real-Time PCR System, PCR and analysis can be performed on one instrument , which is convenient, saves time and reduces costs. Currently, the main application for the High Resolution Melting method is gene scanning, i.e., the identification of heterozygotes in order to discover new variations in target-gene-derived PCR ampli-cons (Figure 1).