Figure 3

Genotyping the 27 bp deletion mutation in healthy Hungarian individuals. Amplification conditions: reaction mixtures contained 200 μM dATP, dCTP, dTTP, and 100 μM dGTP and dITP; 1 μM of each primer (see Methods), 1 ng DNA template, 0.25 U DNA polymerase, 1x reaction buffer, and 1x Q solution in a total volume of 10 μL. Thermocycling conditions: 95°C for 15 min, followed by 35 cycles of 94°C for 1 min; 65°C for 30 sec; 72°C for 1 min, finished by 72°C for 10 min. Electrophoresis: fine resolution agarose gel in 40 mM Tris, 10 mM EDTA pH 8.0 buffer solution at room temperature, 6.6 V/cm electric field, 90 min, followed by 1 μg/mL ethidium bromide staining. The faint diffuse bands in lane 3 migrating slower than the representative fragments presumably contain heteroduplex molecules.