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. Author manuscript; available in PMC: 2009 Aug 1.
Published in final edited form as: Exp Mol Pathol. 2008 Apr 13;85(1):50–58. doi: 10.1016/j.yexmp.2008.03.012

Figure 4.

Figure 4

Combined unlabeled probe and amplicon melting analysis. (A) Asymmetric PCR produces excess copies of strand W and limiting copies of strand C. Excess copies of strand W are available for probe binding, revealing expected and variant sequences under the probe as melting peaks. Sufficient copies of strand C are present so that the full length amplicon duplexes allow variant scanning anywhere within the amplicon. (B) Both product/probe melting transitions and product/product melting transitions are visible on a single negative derivative plot of normalized background corrected fluorescence. Examples of wild-type (green), mutant (red) and heterozygous (blue) melting peaks are illustrated.