Pfu DNA Polymerase

Description

Description:
Pfu DNA Polymerase is a highly thermostable DNA polymerase from hyperthermophilic archaeum Pyrococcus furiosus. The enzyme catalyzes the template-dependent polymerization of nucleotides into duplex DNA in 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 that Pfu has superior thermostability and proof-reading 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.

Applications:

• 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 cloning

Source:
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 8.8
20 mM MgSO₄
100 mM KCl
100 mM (NH₄)₂SO₄
1% Triton
1 mg/ml BSA
2 mM dNTP

Supplied In:
20 mM Tris-HCl (pH 8.0)
40 mM NaCl
0.1 mM EDTA
1 mM DTT
50% (v/v) glycerol

Heat Inactivation:
95% inactive after 1-hour incubation at 98°C

Recommended 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

Additional information

Manual & Protocols

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