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

Citation

Lombal, AJ and Wenner, TJ and Burridge, CP, Assessment of high-resolution melting (HRM) profiles as predictors of microsatellite variation: an example in Providence Petrel (Pterodroma solandri), Genes and Genomics, 37, (12) pp. 977-983. ISSN 1976-9571 (2015) [Refereed Article]

Copyright Statement

Copyright 2015 The Genetics Society of Korea and Springer-Science and Media

DOI: doi:10.1007/s13258-015-0327-9

Abstract

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 (ΔTm) 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 ΔTm, and 25 % (92 vs. 67 %) by using ΔTm 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.

Item Details

Item Type:Refereed Article
Keywords:high-resolution melting analysis, microsatellite, polymorphism, melting temperature range, Pterodroma solandri
Research Division:Biological Sciences
Research Group:Genetics
Research Field:Population, Ecological and Evolutionary Genetics
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Environmental Sciences
Author:Lombal, AJ (Ms Anicee Lombal)
Author:Wenner, TJ (Mr Theodore Wenner)
Author:Burridge, CP (Dr Christopher Burridge)
ID Code:104962
Year Published:2015
Web of Science® Times Cited:1
Deposited By:Zoology
Deposited On:2015-11-30
Last Modified:2017-10-31
Downloads:0

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