Recombinant Pfu DNA Polymerase is purified from an E.coli strain carrying a plasmid with the cloned gene encoding Pyrococcus furiosus DNA polymerase. The enzyme catalyzes the incorporation of nucleotides into duplex DNA in the 5´->3´ direction in the presence of Mg²⁺ at 70-80°C. Pfu DNA Polymerase exhibits 3´-> 5´ exonuclease (proof-reading) activity, but has no detectable 5´->3´ exonuclease activity. The amplified products by Pfu DNA Polymerase can be used for a gene cloning with decreased error rate, and for a site-specific mutagenesis. Pfu DNA Polymerase, like any other polymerases showing proof-reading activity, generates PCR products with blunt end.
Pfu DNA Polymerase is recommended for an amplification of DNA fragment smaller than 7 kbp. Pfu DNA Polymerase is provided with 10x optimized reaction buffer.
Applications
- cDNA amplification for a cloning with high fidelity
- Point mutagenesis
Storage Buffer
- 5 unit/㎕ in 50 mM Tris-HCl, pH 8.2, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol.
Unit Definition
- One unit of enzyme catalyzes the incorporation of 10 nmole of dNTP into a polynucleotide fraction in 30 min at 72°C.
10x Reaction Buffer
- 200 mM Tris-HCl, pH 9.0, 100 mM KCl, 100 mM (NH₄)₂SO₄, 20 mM MgSO₄, 1% Triton X-100, 1 mg/ml BSA.
QC tests
- Activity, SDS-PAGE purity, performance tests, genomic DNA contamination test, confirmation test for the absence of endo and exonucleases.
Storage Condition
- Store at -20℃.
