The mixture was
overlaid with mineral oil, denatured for 5 min at 95 8C and
amplified for 35 cycles (95 8C for 1 min, 55 8C for 1 min, 72 8C
for 1 min) with a final extension of 7 min at 72 8C. The PCR
reactions were carried out in a thermocycler (PTC-100, MJ
Researcher, USA).