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. Author manuscript; available in PMC: 2014 Jul 11.
Published in final edited form as: Nat Protoc. 2012 Feb 23;7(3):542–561. doi: 10.1038/nprot.2012.005
Component Volume Final concentration
5 X High-Fidelity buffer 10 µl 1 X
dNTPs (2.5 mM each) 4 µl 0.2 mM
100 µM PCR Forward
primer		 0.5 µl 1 µM
100 µM PCR Reverse
primer		 0.5 µl 1 µM
Phusion polymerase (2 U µl−1) 0.5 µl 1 U
cDNA with magnetic beads from Step 50 0.5 – 2 µl -
Water variable -
Total volume 50 µl -