Pfu DNA Polymerase

Description

• Pfu DNA Polymeraseis a thermostable enzyme isolated from hyperthermophilic archaea Pyroccocus sp. (1).
• Ultrapure recombinant enzyme.
• Unmodified enzyme replicates DNA at 74°C and exhibits over 95% activity after 1-hour incubation at 95°C.
• The enzyme catalyzes the polymerization of nucleotides into duplex DNA in the 5´->3´ direction in the presence of magnesium ions.
• The enzyme exhibits the 3´->5´ proofreading activity, resulting in over 10-fold higher PCR fidelity than possible with Taq DNA Polymerases (2).
• Pfu DNA Polymerase is recommended for use in high-fidelity PCR, PCR of GC-rich sequences or problematic secondary structures and cloning of blunt-ended PCR products.

Unit Definition

One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmoles of dNTP into acid-insoluble material in 30 min at 74°C.

Reaction Conditions

50 mM Tris-HCl (pH 9.0 at 25°C)
50 mM NaCl, 5 mM MgCl₂
200 μM each of dATP, dCTP, dGTP, dTTP (a mix of unlabeled and [³H]dTTP)
10 μg activated calf thymus DNA
0.1 mg/ml BSA
Final volume 50 μl

Quality Control

All preparations are assayed for contaminating endonuclease, 3'-exonuclease, and nonspecific single- and double-stranded DNase activities. Typical preparations are greater than 95% pure, as judged by SDS polyacrylamide gel electrophoresis.