Pfu DNA polymerase (with dNTPs)

Description

Pyrococcus furiosus DNA polymerase (Pfu DNA polymerase) gene was expressed in E. coli in large quantities and highly purified. The enzyme has thermostable DNA polymerase activity and 3’ →5’ exonuclease (proofreading) activity. The MW is 90 kDa, same as that of the natural Pfu DNA polymerase.

• Pfu DNA polymerase is thermostabe and has low error rates.
• It is suitable for PCR and primer extension reactions that require high fidelity synthesis.
• Pfu DNA polymerase-generated PCR fragments are blunt-ended.

Applications

1. Cloning
2. DNA expression
3. site-directed mutagenesis

Specification

Storage Conditions: 50mM Tris-HCl (pH 8.2), 0.1mM EDTA, 1mM DTT, 50% glycerol, 0.1% Tween20, 0.1% Igepal CA-630

Store at -20°C

Concentration: 2.5 units/ul, where one unit is defined as the amount of enzyme that can incorporate 10 nmols of dNTPs into an acid-insoluble material in 30 minutes at 72°C when activated salmon sperm DNA was used as template/primer.

Quality Assurance:

Greater than 95% of protein determined by SDS-PAGE (CBB staining) (Fig.1)

The absence of endonucleases and exonucleases was confirmed.

PCR Test: Good amplification result was obtained in PCR reaction using λDNA as a template (Fig. 2).

Reagents Supplied with Enzyme:

10 x Reaction Buffer (Pfu): 200mM Tris-HCl (pH 8.8), 100mM KCl, 100mM (NH₄)₂SO₄, 20mM MgSO₄, 1% TritonX-100, 1 mg/ml BSA

2.5mM (each) dNTPs

Fig. 1 SDS-PAGE of Pfu DNA polymerase

Fig. 2 Amplification of λ DNA