1 min DNA
denaturation step at 94 ºC, 2 min annealing
step at 36 ºC, 2 min extension step at 72 ºC
and last cycle at 72 ºC for 7 min, and an
optional soak period at 4 ºC. Amplification
products were loaded on 1% agarose gel
and stained with ethidium bromide (0.5
mg/ml