I'fD AppUcations in Genetics
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Fig.4A-C. Rapid cycle PCR and genotyping by melting point analysis of a methionine homozygous sample using probe set 1 and different amounts of reverse primer 1. Amplification contained 5 pmol of forward primer. The numbered curves and lanes show results obtained in the
presence of these amounts of reverse primer: 1 (5 pmol); 2 (2.5 pmol); 3 (1 pmol); 4 (0.5 pmol).
A Fluorescence signal detected at the annealing step of each cycle during the amplification.
B Melting point analysis of the PCR products. Melting peaks between 57°C and 59°C. C Agarose
gel electrophoresis of PCR products removed from the glass capillaries. M, 100-bp molecular
weight marker; -d(F2)/dT, negative derivative of fluorescence with respect to temperature