PCR was performed as follows: Initial denaturation of template DNA was carried out at 94 ºC for 5 min, then denaturation at 94 ºC for 1 min, annealing at 55 ºC for 1 min and elongation for 3 min at 72 ºC for 35 cycles followed by final elongation at 72 ºC for 10 min.