GT-PFu DNA Polymerase is a recombinant form of highly thermostable DNA polymerase from the hyperthermophilic archaeum Pyrococcus furiosus which is expressed in E.Coli. The enzyme catalyzes the template-dependent polymerisation of nucleotides into duplex DNA in the 5’→3′ direction. It exhibits 3’→5′ exonuclease (proofreading) activity that enables the polymerase to correct nucleotide incorporation errors. It has no 5’→3′ exonuclease activity. Recombinant changes in enzyme make it possible to facilitate faster polymerization.
The error rate of PFu DNA Polymerase in PCR is 2.6 x 10-6 errors per nucleoid per cycle as determined by a modified method described by Lundberg et al. 1991. Accordingly the accuracy of PCR is 3.8 x 10 -5. This enzyme has no detectable reverse transcriptase activity. dUTP, dITP, and primers containing these nucleotides should not be used in PCR with PFu DNA Polymerase because the binding of this enzyme to DNA templates with uracil and hypoxanthine stalls DNA synthesis.
Features & Benefits:
- Eight times more accurate than Taq DNA polymerase
- Highly thermostable—remains 95% active after 2 hours incubation at 95°C
- Generates blunt-end PCR products
- Incorporates modified nucleotides (e.g., biotin-, digoxigenin-, fluorescently-labeled nucleotides)
- Increased PCR yields with PR DNA Polymerase
- Can be used in DNA cloning and gene synthesis
- High fidelity PCR
- Generation of PCR products for cloning and expression
- RT-PCR for cDNA cloning and expression
- Blunt-end PCR cloning