Abstract
High-resolution melting (HRM) analysis is a simple, closed tube, post-PCR method used to identify genetic variation. The method is highly sensitive and can discriminate DNA sequence variants based on length (such as insertions or deletions), composition (such as single nucleotide polymorphisms, i.e., SNP) or strand complementarity (such as heterozygous or homozygous material). The technique involves PCR amplification of a target sequence in the presence of a fluorescent double-stranded DNA (dsDNA) binding dye, melting of the fluorescent amplicons, and subsequent interpretation of melt curve profiles. Here, we describe general considerations for assay design, PCR amplification, and HRM analysis.
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Tucker, E.J., Huynh, B.L. (2014). Genotyping by High-Resolution Melting Analysis. In: Fleury, D., Whitford, R. (eds) Crop Breeding. Methods in Molecular Biology, vol 1145. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0446-4_5
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DOI: https://doi.org/10.1007/978-1-4939-0446-4_5
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-0446-4
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