|Source||Isolated from E.coli strain that carries recombinant plasmid containing the modified Pfu DNA Polymerase gene.|
|Description||PfuSE DNA Polymerase is improved (more stable) variant of Pfu DNA Polymerase. The enzyme catalyzes the DNA-dependent polymerization of nucleotides into DNA duplex in the 5`- 3` direction in the presence of magnesium ions. PfuSE DNA Polymerase possesses a 3`- 5` exonuclease (proof-reading) activity, that allows the polymerase to correct the nucleotide incorporation errors.|
The enzyme produces DNA-products with blunt ends.
- PfuSE DNA Polymerase is useful for high fidelity
synthesis and the end polishing.
|Unit definition||One unit of enzyme catalyzes the incorporation of 10 nanomoles of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE- 81) in 30 min at 72°C.|
Unit Assay Conditions: 1 x PfuSE-DNA-polymerase buffer, 0.1 mg/ml BSA,
200 mg/ml activated calf thymus DNA,
0.2 mM of each dNTP, 0.4 MBq/ml [3 H]- dTTP in 50 ml reaction mix.
|Reaction buffer||SE-buffer PfuSE 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||1. Do not use dU-contained templates.|
2. This enzyme is not recommended for the experiments with very low-annealing temperature amplification approaches (e.g. RAPDs, Random Amplified Polymorphic DNAs).
3. Prepare PCR mix at 0°C and place it in the
amplificator preheated to 95°C.
Recommended enzyme volumes to be added to the reaction mix: We recommend using 2.5 enzyme units (0.5 μl) in a final reaction volume of 50 μl.
|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.