Pfu/Psp DNA Polymerase

Features:
Pfu/Psp DNA polymerase replicates DNA at 75°C catalyzing the polymerization of nucleotides into duplex DNA in the 5´=>3´ direction in the presence of Mg⁺. Pfu DNA polymerase possesses 3′ to 5′ exonuclease proof reading activity that enables the polymerase to correct nucleotide-misincorporation errors. The enzyme has no 5’=>3′ exonuclease activity.

Applications:
– blunt end PCR cloning
– PCR and primer extension where “high fidelity” is required
– Site-directed mutagenesis

Description:
Pfu/Psp DNA polymerase, isolated from the archae bacteria Pyrococcus furiosus/species is a thermostable Polymerase of approximately 90000 daltons.  Base misinsertions that may occur during polymerization are rapidly excised by the proofreading activity of the polymerase. The Pfu DNA Polymerase has no detectable reverse transcriptase activity.

Concentration: 5 u/µl

Unit definition:
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nM of dNTPs into acid insoluble material in 30 minutes at 75°C.

Storage Buffer:
50 mM Tris-HCl, pH 8.2, 0.1 mM EDTA, 0.1% Tween 20, 0.1% Nonident P40, 1 mM DTT, 50% Glycerol

Reaction Buffer 10 X:
100 mM KCl, 160 mM (NH₄)₂SO₄, 20 mM MgSO₄, 200 mM Tris-HCl, pH8.8, 1% Triton X-100, 1 mg/ml BSA

Usage:
Standard protocol:
– Do not use dUTP or dITP or primers containing these nucleotides