Genotyping of the Methionine-Valine Polymorphism at Codon 129 of the Human Prion Protein 1m
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Fig.3A-C. Rapid cycle PCR and genotyping by melting point analysis of a heterozygous sample
using probe set 1 and different amounts of reverse primer 1. Each 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; negative control without DNA); 2 (5 pmol); 3 (2.5 pmol);
4 (1 pmol); 5 (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 and approximately 67-68°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