Genotyping of the Methlonine-Valine Polymorphism at Codon 129 of the Human Prion Protein 1m
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Fig. 5. Rapid cycle PCR and genotyping by melting point analysis of a valine 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 (I 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 peak approximately 67°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
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Temperature (0
M
M
c
-
18Sbp
Fig. 5. Rapid cycle PCR and genotyping by melting point analysis of a valine 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 (I 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 peak approximately 67°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