cDNA and 4.5ฮผl ddH2O in a total volume of 20μl. The PCR reactions were carried out and the cycling program was set as follows: an initial denaturing step at 94°C for 4min, followed by 35 cycles of 94°C for 15s, annealing at 52-64°C for 20s, and extension at 72ยฐC for 20s, and a final