2.4. PCR conditions
The PCR mixture contained 1 PCR buffer, 200 lM dNTPs, 0.6
units of Taq Polymerase, 20 pmol each of the sense and antisense
primer and 75 ng of template DNA. The following thermal cycle
conditions were used: denaturation of DNA at 95 C for 5 min, followed
by 30 cycles of denaturation at 95 C for 30 s, annealing at
65 C for 30 s and, extension at 72 C for 30 s. A final extension step
was performed at 72 C for 5 min. Amplification products were
separated by agarose gel-electrophoresis using a 2% (w/v) agarose
gel in 1 TAE, stained with ethidium bromide solution and visualized
with a BioRad Gel Doc.