Component	Amount (μl)	Final concentration
Ammonium PCR buffer, 10×	2.5	1×
dNTP, 100 mM (25 mM each)	0.25	1 mM
QCgFwd primer, 25μM	0.25	0.25 μM
QCgRev primer, 25μM	0.25	0.25 μM
QCgX primer, 2.5μM	0.25	0.025 μM
TEMPase Hot Start DNA polymerase	0.25	1.25 U
DNA template from Step 2, 50 pg μl−1	1.0	50 pg
ddH2O	20.25
Total	25