Pfu DNA Polymerase (native)

Detailní popis

Thermo Scientific 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.

Highlights

• 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 native Pfu DNA Polymerase

Applications

• High fidelity PCR
• Generation of PCR products for cloning and expression
• RT-PCR for cDNA cloning and expression
• Blunt-end PCR cloning (see Reference 1)

Includes

• Pfu DNA Polymerase (native or recombinant, 2.5 U/µL)
• 10X Pfu Buffer with MgSO₄
• 10X Pfu Buffer
• 25 mM MgSO₄

Note

The error rate of Pfu DNA Polymerase in PCR is 2.6 x 10^-6 errors per nt per cycle as determined by a modified method described by Lundberg et al. (see Reference 2). Accordingly, the accuracy of PCR is 3.8 x 10⁵.  Accuracy is an inverse of the error rate and shows an average number of correct nucleotides incorporated before an error occurs. The Pfu DNA Polymerase 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 (see References 3,4).