Pfu DNA Polymerase is a thermostable enzyme that replicates DNA at 75°C. It catalyzes the polymerization of nucleotides into duplex DNA in the 5´→3´ direction in the presence of magnesium. The enzyme has a molecular weight of approximately 90,000 daltons as estimated from the predicted amino acid sequence and exhibits 3´→5´ exonuclease (proofreading) activity. Pfu DNA Polymerase is recommended for use in PCR and primer extension reactions that require high fidelity.
Storage Solution : 5 U/ul Pfu DNA Polymerase, 20 mM Tris-HCl, 0.1 mM EDTA, 0.1% Tween20, 0.1% triton X100, 1 mM DTT, 100 mM KCl, 50%Glycerol、pH 8.2 @ 25°C
10* Reaction Buffer: 200 mM Tris-HCl (pH 8.8) , 20 mM MgSO4, 100 mM KCl, 100 mM (NH4) 2SO4, 1% Triton X-100, 1 mg/ml nuclease-free BSA
Note: 1. It is critical to withhold Pfu DNA Polymerase until after the addition of dNTPs; otherwise, the proofreading activity of the polymerase may degrade the primers, resulting in nonspecific amplification and reduced product yield.
2. Assemble on ice.
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 74°C
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