4.Load tubes to iCycler PCR machine and perform PCR
amplification by following cycle conditions: initial activation at
94 °C for 5 min, then 35 cycles at 94 °C for 30 s, 60 °C for
1 min, and 72 °C for 1 min 30 s, and afinal elongation at
72 °C for 10 min. The hold temperature is 4 °C.