Reagent	Volume per reaction	Volume for N reactions	Final concentration
Water	6.2 μL	6.2 × N μL	–
5× TaqMan EZ buffer	5 μL	5 × N μL	1×
Manganese acetate, 25 mM	3 μL	3 × N μL	3.0 mM
dATP, 10 mM	0.75 μL	0.75 × N μL	0.3 mM
dUTP, 10 mM	1.5 μL	1.5 × N μL	0.6 mM
dCTP, 10 mM	0.75 μL	0.75 × N μL	0.3 mM
dGTP, 10 mM	0.75 μL	0.75 × N μL	0.3 mM
Forward primers, 50 μM	0.4 μL	0.4 × N μL	0.8 μM
Reverse primers, 50 μM	0.4 μL	0.4 × N μL	0.8 μM
Probe, 10 μM	1 μL	1 × N μL	0.4 μM
rTth DNA polymerase (2.5 U/μL)	1 μL	1 × N μL	2.5 U/reaction
AmpErase UNG (1 U/μL)	0.25 μL	0.25 × N μL	0.25 U/reaction
Total	21 μL	21 × N μL	–