f-Pfu DNA Polymerase

Description

f-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. f-Pfu DNA Polymerase exhibits 3'→5' exonuclease (proofreading) activity that enables the polymerase to correct nucleotide-misincorporation errors, 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.

The extension rate of f-Pfu DNA Polymerase is about 600 bp/min in standard condition. The appropriate reaction temperature is 65~75℃, the work concentration of dNTPs is 100~300μM, the work concentration of Mg²⁺ is 2~3mM, and the suitable 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.

Protocol.pdf