Assessment of high-resolution melting (HRM) profiles as predictors of microsatellite variation: an example in Providence Petrel (Pterodroma solandri)

作者：Lombal Anicee J^*; Wenner Theodore J; Burridge Christopher P

来源：Genes & Genomics, 2015, 37(12): 977-983.

DOI：10.1007/s13258-015-0327-9

• High-resolution melting analysis
• Microsatellite
• Polymorphism
• Melting temperature range
• Pterodroma solandri

High-resolution melting (HRM) analysis is an emerging technology to screen microsatellites for polymorphism. A potential issue surrounding this method is that amplicon sizes for HRM should typically be short (80-100 bp) for highest sensitivity to reveal polymorphism via the presence of two peaks in the curve of the derivative of fluorescence over temperature (dF/dT). In contrast, microsatellite amplicons are typically 100-400 bp. Therefore, we compared HRM analysis melting temperature range (Delta T-m) and multiple dF/dT peaks for predicting microsatellite polymorphism. We assessed polymorphism at 27 microsatellite loci, with estimated lengths of 122-321 bp, in Providence Petrel (Pterodroma solandri). We validated HRM assessment using traditional capillary electrophoresis (CE). While 100 % of loci exhibiting multiple peaks in the dF/dT curve were confirmed as polymorphic by CE, 16 % improvement in sensitivity (83 vs. 67 %) was achieved by using Delta T-m, and 25 % (92 vs. 67 %) by using Delta T-m in addition to multiple dF/dT peaks. We suggest HRM melting temperature range as new predictor of polymorphism that can be used to rapidly assess microsatellites polymorphism.

• 出版日期2015-12