The thermal cycling included an initial denaturation
step at 95 °C for 10 min; 30 cycles of a denaturation step at 95 °C for
30 s, an annealing step at 50 °C for 1 min, an extension step at 72 °C
for 2 min; and a final extension at 72 °C for 10 min. The PCR product
was checked using 0.8% (w/v) agarose gel electrophoresis. The gel was
visualized using an Image Master® VDS.