HIGH-RESOLUTION DNA MELTING ANALYSIS: ADVANCEMENTS AND LIMITATIONS

摘要: Recent advances in fluorescent dyes, methods, instruments and software for DNA melting analysis have created versatile new tools variant scanning genotyping. High resolution (HRM or HRMA) is faster, simpler, less expensive than alternative approaches requiring separations labeled probes. With the addition of a saturating dye before PCR followed by rapid products, sensitivity heterozygote 100%. Specificity can be increased identifying common polymorphisms with small amplicon melting, unlabeled probes snapback primers to decrease sequencing burden. However, some homozygotes require mixing identification. Furthermore, different heterozygotes may produce curves so similar each other that, although they clearly vary from homozygous variants, are not differentiated other. Nevertheless, experimental return minimal effort great. This focus issue Human Mutation includes concise, timely review on high comparison denaturing gradient gel electrophoresis, integration qPCR copy number assessment, combined probe genotyping single curve, applications mitochondrial genome BRCA1. Hum Mutat 30, 857–859, 2009. © 2009 Wiley-Liss, Inc.