Quantitative high resolution melting: two methods to determine SNP allele frequencies from pooled samples

Capper, Roxana L., Jin, Young K., Lundgren, Petra B., Peplow, Lesa M., Matz, Mikhail V., and van Oppen, Madeleine J.H. (2015) Quantitative high resolution melting: two methods to determine SNP allele frequencies from pooled samples. BMC Genetics, 16. 62. pp. 1-13.

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Abstract

Background: The advent of next-generation sequencing has brought about an explosion of single nucleotide polymorphism (SNP) data in non-model organisms; however, profiling these SNPs across multiple natural populations still requires substantial time and resources.

Results: Here, we introduce two cost-efficient quantitative High Resolution Melting (qHRM) methods for measuring allele frequencies at known SNP loci in pooled DNA samples: the "peaks" method, which can be applied to large numbers of SNPs, and the "curves" method, which is more labor intensive but also slightly more accurate. Using the reef-building coral Acropora millepora, we show that both qHRM methods can recover the allele proportions from mixtures prepared using two or more individuals of known genotype. We further demonstrate advantages of each method over previously published methods; specifically, the "peaks" method can be rapidly scaled to screen several hundred SNPs at once, whereas the "curves" method is better suited for smaller numbers of SNPs.

Conclusions: Compared to genotyping individual samples, these methods can save considerable effort and genotyping costs when relatively few candidate SNPs must be profiled across a large number of populations. One of the main applications of this method could be validation of SNPs of interest identified in population genomic studies.

Item ID: 41726
Item Type: Article (Research - C1)
ISSN: 1471-2156
Keywords: pooled genotyping, allele frequency, high resolution melting, SNP, population
Additional Information:

© Capper et al. 2015. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Funders: Australian Institute of Marine Science (AIMS), National Science Foundation (NSF)
Projects and Grants: NSF DEB-1054766
Date Deposited: 08 Dec 2015 18:06
FoR Codes: 06 BIOLOGICAL SCIENCES > 0604 Genetics > 060499 Genetics not elsewhere classified @ 50%
06 BIOLOGICAL SCIENCES > 0602 Ecology > 060205 Marine and Estuarine Ecology (incl Marine Ichthyology) @ 50%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100%
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