|Source||Isolated from E.coli strain that carries the cloned DNA polymerase gene from Pyrococcus furiosus.|
|Description||Pfu DNA Polymerase catalyzes the DNA-dependent polymerization of nucleotides into duplex DNA in the 5’ -> 3’ direction in the presence of magnesium ions. The enzyme also exhibits 3’ -> 5’ exonuclease (proofreading) activity, that enables the polymerase to correct nucleotide incorporation errors. Products of reaction have blunt ends.|
Pfu DNA Polymerase useful for high fidelity synthesis and polishing ends.
|Unit definition||One unit of enzyme catalyzes the incorporation of 10 nanomoles of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE- 81) in 30 minutes at 72°C.
Unit Assay Conditions: 1 x Pfu DNA polymerase buffer,
0.1 mg/ml BSA, 200μg/ml activated calf thymus DNA,
0.2 mM of each dNTP,
0.4MBq/ ml [3H]- dTTP in 50 μl reaction mix. |
|Reaction buffer||SE-buffer Pfu DNA polymerase, (20 mM Tris-HCl (pH 8.8 at 25°C); 10 mM KCl, 10 mM (NH4)2SO4, 2 mM MgSO4, 0.1 % Triton X-100 )+ 100 μg/ml BSA)|
|Storage conditions||10 mM K2HPO4 (pH 7.4); 0.1 mM DTT; 0.1 mM EDTA; 0.5 % Tween 20; 50% glycerol. Store at -20°C.|
|Notes||To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.|
The recommended amount of enzyme
is 2.5 u per 50μl of a total reaction volume.
|Quality control||The enzyme is purified free of contaminating endonucleases and
Some applications in which this product can be used may be covered by patents issued and applicable in the United States, European Union and certain countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a patent license depending upon the particular application in which the product is used.
|MSDS:||Download MSDS as PDF|