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
- Cloning
- DNA expression
- 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
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