Advances in Nucleic Acid Detection and Quantification 433 Mutation scanning using high-resolution melting

Mutation scanning techniques are used to detect sequence variants without the need for prior knowledge of the identity or precise location of the variant, in contrast with genotyping techniques, which determine the status of a specific variant. High-resolution melting is a recently developed method that shows great potential as a mutation scanning technique. Sensitivity and specificity for mutation detection are extremely high and the technique also has advantages of cost and throughput. Practical considerations for successful mutation scanning by high-resolution melting are also discussed in this review. Principle of HRM (high-resolution melting) HRM is a simple, PCR-based method for detecting DNA sequence variation by measuring changes in the melting of a DNA duplex. Melting of double-stranded DNA molecules is influenced by several factors. Some of these, such as the length,GC content and sequence, are properties of the indivi-dual molecule [1]. Others are not: these include the ionic strength of the buffer solution, the DNA concentration an