Pfu DNA Polymerase is a highly thermostable DNA polymerase from the hyperthermophilic archaeum Pyrococcus furiosus.
Description:Pfu DNA Polymerase is a highly thermostable DNA polymerase from the hyperthermophilic archaeum Pyrococcus furiosus. The enzyme catalyzes the template-dependent polymerization of nucleotides into duplex DNA in the 5\'->3\' direction. Pfu DNA Polymerase also exhibits 3\'->5\' exonuclease (proofreading) activity, that enables the polymerase to correct nucleotide incorporation errors. It has no 5\'->3\' exonuclease activity. The main difference between Pfu and alternative enzymes is Pfu\'s superior thermostability and \'proofreading\' properties. Unlike Taq DNA polymerase, Pfu DNA polymerase also possesses 3\'->5\' exonuclease proofreading activity, resulting in PCR fragments with fewer errors than Taq-generated PCR inserts. Pfu DNA polymerase is efficient for techniques that require high-fidelity DNA synthesis, but can also be used in conjunction with Taq polymerase to obtain the fidelity of Pfu with the speed of Taq polymerase activity.Application:- High-fidelity PCR and primer-extension reactions- Generation of PCR products for cloning and expression- PCR cloning and blunt-end amplification product generation- RT-PCR for cDNA cloning and expression- Site-directed mutagenesis- Blunt-end PCR cloningSource:Thermostable DNA polymerase from hyperthermophilic archaeon pyrococcus furiosus.Unit Definition:One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dNTPs into acid insoluble material in 30 minutes at 74°C under standard DNA polymerase assay conditions.Supplied With:10x Pfu Reaction Buffer (with dNTPs) 10x Pfu Reaction Buffer (with dNTPs) 200mM Tn3 pH 8.820 mM MgSO4100 mM KCl100 mM (NH4)2SO41% Triton1 mg/ml BSA2 mM dNTP Supplied In: 20 mM Tris-HCl (pH 8.0)40 mM NaCl0.1 mM EDTA1 mM DTT50% (v/v) glycerolHeat Inactivation:95% inactive after 1-hour incubation at 98°CRecommended Reaction Conditions:1X Pfu buffer, 200 µM each dNTP, 0.1-0.5µM each primer, 5 units Pfu DNA polymerase enzyme, 1-100ng plasmid template DNA, or 100-250ng genomic template DNA.Recommended Storage Condition: -20ºC |
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