By

Dr Michael Tavaria

Scientific Applications Support Manager

Abstract:

High Resolution Melt (HRM) analysis is a relatively new technique that enables small differences in DNA sequence to be identified quickly, easily and cheaply. HRM is based on post-PCR melting curve techniques that have existed for quite some time, but it is enabled by the recent availability of improved double-stranded DNA binding dyes, advanced real time PCR instruments, reagents and analysis software. HRM can be used for many applications such as SNP genotyping, mutation screening, methylation analysis and viral/bacterial strain identification. The HRM workflow is fairly simple, but success requires considerable care in PCR primer design and validation, reagent selection, DNA preparation and analysis methods. Discussion of these important topics should provide potential and existing users with the knowledge required to optimise their use of the HRM technique.