Reagent	Final Concentration	Volume (μL)
HotStarTaq DNA Polymerase (5 U/μL) (Cat#203209, QIAGEN)	0.05 U/μL	42
Forward Primer Mixture (50 μM) (see Note above)	0.29 μM	23
Reverse Primer (50 μM) (see Note above)	0.19 μM	15
dNTP (25 mM) (see Note blow)	0.31 mM	48
10× PCR Buffer (supplied with HotStarTaq DNA Polymerase)	~1.1×	420
40% Sucrose Loading Buffer (see above)	8.2%	800
Nuclease-Free Water (Cat#129114, QIAGEN)	n/a	2,552
Total	n/a	3,900