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Website: http://www.roche.applied.science.com
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Melting Curve-based Analysis Of Adjacent SNPs
by Thomas Froehlich and Michael Hoffmann
Introduction
Melting curve analysis (MCA) is a robust and well-established method to detect mutations or single nucleotide polymorphisms (SNPs) in amplicons generated by Real-Time PCR.(1, 2)
Due to its high-end optical and thermal characteristics, the LightCycler 480 System (Roche Applied Science, Indianapolis, IN, Figure 1) is suited for this application. Known SNPs present in PCR amplicons can be investigated with sequence-specific, labeled probes that bind with different strength to the different alleles or allele combinations in an SNP-containing region, depending on whether they fully match the target sequence or contain mismatches.
Due to the equations in this article, we have made a PDF
available for you to download.
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