Table 5. Thermal cycling program for the second round of amplification.

Step	Temperature	Time	Cycles
Initial denaturation	94°C	2 min	1
Denaturation	94°C	30 s	40
Annealing	55°C	30 s	40
Extension	72°C	20 s	40
Final extension	72°C	10 min	1
Hold	16°C	∞	-