Reagents	Volume (μl)
Ligation product	39.8
10× high fidelity PCR buffer	5
50 mM MgSO4	2
dNTPs (10 mM each)	1
10 μM RP1	1
10 μM 3′ index primer	1
Platinum® Taq DNA polymerase high fidelity	0.2
Total	50