Component	Amount (μl)	Final concentration
Herculase II PCR buffer, 5×	10	1×
dNTP, 100 mM (25 mM each)	1	2 mM
Fwd primer (10 μM)	1	0.2 μM
Rev primer (10 μM)	1	0.2 μM
Herculase II fusion polymerase	1
MgCl2 (25 mM)	2	1 mM
DNA template	1	2 ng μl−1
ddH2O	33
Total	50