Reagent	Volume per reaction	Volume for N reactions	Final concentration
10× PCR buffer	5 μL	5 × N μL	1×
MgCl2, 25 mM	5 μL	5 × N μL	2.5 mM
dNTP, 10 mM	0.5 μL	0.5 × N μL	0.1 mM
Forward primers, 10 μM	1.25 μL	1.25 × N μL	0.25 μM
Reverse primers, 10 μM	1.25 μL	1.25 × N μL	0.25 μM
DNA polymerase (5 U/μL)	0.25 μL	0.25 × N μL	1.25 U/reaction
Water	34.75 μL	34.75 × N μL	−
Total	48 μL	48 × N μL	−