Step

Temperature

Time

Number of Cycles

Initial Denaturation

95˚C

2 minutes

1 cycle

Denaturation

95˚C

0.5 - 1 minute

25 - 35 cycles

Annealing

42˚C - 65˚C*

0.5 - 1 minute

25 - 35 cycles

Extension

72˚C

1 minutes/kb

25 - 35 cycles

Final Extension

72˚C

5 minutes

1 cycle

Soak

4˚C

indefinite

1 cycle