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. Author manuscript; available in PMC: 2018 Feb 1.
Published in final edited form as: Nat Protoc. 2017 Jul 20;12(8):1659–1672. doi: 10.1038/nprot.2017.055
Component Amount per
reaction (µL)		 Final
amount/concentration
Ligated DNA library (0.5 ng/uL) 10 5ng
Water 2.6 -
5× Phusion HF Buffer 4 1×
10 mM dNTPs 1 500 µM
Primer A (25 µM) 0.4 0.5 µM
Primer B (25 µM) 0.4 0.5 µM
Phusion DNA Polymerase (2000 U/mL) 0.4 2 U
10× SYBR Green I 1.2 0.6×