Pfu DNA Polymerase

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 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