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The reaction mixture had 100 ng of template DNA, 200 nM of forward and reverse primers, 2x of Phusion mixture (40 mM TRIS pH 8.8, 4 mM MgCl 2 , 120 mM KCl, 20 mM (NH 4 ) 2 SO 4 , 0.02 mM EDTA, 0.2% TritonX-100, 8% glycerol, 0.005% Xylene Cyanol FF, 0.05% Orange G, 0.4 mM dNTPs, 0.04 U/L Phu-Sso7d Polymerase) and nuclease-free water to a total volume of 50 l

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