Cycle	cDNA synthesis	Denaturation	Annealing	Extension	Hold
1	50°C for 30 minutes
2		94°C for 2 minutes
3–47		94°C for 15 seconds	50°C for 30 seconds	68°C for 1 minute
48				1 cycle: 68°C for 5 minutes.
49					4°C for up to 1 week