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. Author manuscript; available in PMC: 2015 May 1.
Published in final edited form as: Methods Mol Biol. 2015;1239:105–132. doi: 10.1007/978-1-4939-1862-1_6
 5 × Phusion HF Buffer 10 μl
 10 mM dNTPs mix 1 μl
 Phusion High-Fidelity DNA Polymerase 0.5 μl
 Gel-extracted PCR fragments 1.5 μl each
 10 pmol/μl the ‘Up2’ and ‘Down2’
primers		 2.5 μl each
 Water Added to make a final volume of 50 μl