High-resolution DNA melting analysis for simple and efficient molecular diagnostics.
High-resolution melting of DNA is a simple solution for genotyping, mutation scanning and sequence matching. The melting profile of a PCR product depends on its GC content, length, sequence and heterozygosity and is best monitored with saturating dyes that fluoresce in the presence of double-stranded DNA. Genotyping of most variants is possible by the melting temperature of the PCR products, while all variants can be genotyped with unlabeled probes. Mutation scanning and sequence matching depend on sequence differences that result in heteroduplexes that change the shape of the melting curve. High-resolution DNA melting has several advantages over other genotyping and scanning methods, including an inexpensive closed tube format that is homogenous, accurate and rapid. Owing to its simplicity and speed, the method is a good fit for personalized medicine as a rapid, inexpensive method to predict therapeutic response.
Reed
Gudrun H
GH
Department of Pathology, University of Utah Medical Center, 5B418, 50 North Medical Drive, Salt Lake City, UT 84132, USA.
Kent
Jana O
JO
Wittwer
Carl T
CT
D000818:Animals; D004252:DNA Mutational Analysis / Q000379:methods* / Q000639:trends; D006358:Hot Temperature; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods* / Q000639:trends; D009691:Nucleic Acid Denaturation / Q000235:genetics*