Pfu DNA Polymerase (500 units)

Pfu DNA Polymerase is a thermostable enzyme with a molecular weight of 90 kDa. It catalyzes the polymerization of nucleotides into duplex DNA in the 5'→3' direction, resulting in blunt-ended PCR products without 3'-dA overhangs. Pfu DNA Polymerase exhibits 3'→5' exonuclease (proofreading) activity that enables the polymerase to correct the mis-incorporation of nucleotide, and lacks 5'→3' exonuclease activity. It is suitable for PCR and primer extension reaction that requires high fidelity when the PCR fragment is relatively shorter than 3kb.

The extension rate of Pfu DNA Polymerase is about 600bp/min in standard condition. The appropriate reaction temperature is 65ºC~75ºC, the working concentration of dNTPs is 100-300μM, the working concentration of Mg2+ is 2~3mM, and the optimal pH is 8.1~9.1. The amount of enzyme is 1~1.5unit for 20μl PCR reaction, while 2~3units for 50μl PCR reaction.

Reaction Buffer (PCR Buffer, 10X):

100mM KCl, 160mM (NH4)2SO4, 20mM MgSO4, 200mM Tris-HCl (pH8.8), 1% Triton X-100, 1mg/ml BSA

Storage Buffer

Pfu DNA Polymerase is supplied in 50 mM Tris-HCl (pH 8.2), 0.1 mM EDTA, 0.1% Tween20, 0.1% NP-40, 1mM DTT, and 50% Glycerol.

Unit Definition