DNA Polymerase, Pfu, Recombinant
|Cloning||Storage: -20°CShipping: Dry Ice|
Recombinant Pfu DNA Polymerase (Pfu) has been shown to exhibit superior thermostability and proofreading properties compared to other thermostable polymerase. Unlike Taq DNA Polymerase, highly thermostable Pfu possesses 3' to 5' exonuclease proofreading activity that enables the polymerase to correct nucleotide-misincorporation errors. This means that Pfu generated PCR fragments will have fewer errors than Taq-generated PCR inserts. The error rate for Pfu is reported to be 7 to 10 fold lower than that of nonproofreading Taq DNA polymerase, and 2 to 30 fold lower than other proofreading enzymes. Using Pfu in your PCR reactions results in blunt-ended PCR products, which are ideal for cloning into blunt-ended vectors. Pfu is superior for techniques that require high-fidelity DNA synthesis.
E.coli strain carrying a plasmid with the cloned gene encoding the hyperthermophilic archae Pyrococcus furiosus DNA Polymerase.
1. Ideal for high-fidelity amplification
2. 3'-5' exonuclease activity provides a low error rate.
3. One of the most thermostable DNA polymerases known.
4. Lack of extendase activity means no unwanted 3’ overhangs.
5. Optimal for blunt-end PCR cloning.
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10nmoles of dNTPs into acid insoluble material in 30 minutes at 74°C under standard DNA polymerase assay conditions.
D3935-65B110X Reaction Buffer:
Supplied as a liquid as 200mM Tris-HCl, pH 8.75, 100mM potassium chloride, 20mM magnesium sulfate, 100mM ammonium sulfate, 1% Triton X-100, 1mg/ml BSA.
Recomended Dilution Buffer:
When diluted 1:10, this buffer contains 20mM Tris-HCl, pH 8.75, 10mM potassium chloride, 2mM magnesium sulfate, 10mM ammonium sulfate, 0.1% Triton X-100, 0.1mg/ml BSA.
Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
E.coli strain carrying a plasmid with the cloned gene encoding the hyperthermophilic archae Pyrococcus furiosus DNA Polymerase
EC No: 184.108.40.206
Accepted Name: DNA-directed DNA polymerase
Reactions: deoxynucleoside triphosphate + DNAn = diphosphate + DNAn+1
Alternate Names: DNA polymerase II subunit A; DNA polymerase epsilon, catalytic subunit A; DNA polymerase II; DNA polymerase ; DNA polymerase ; DNA polymerase ; DNA nucleotidyltransferase (DNA-directed); DNA nucleotidyltransferase (DNA-directed); deoxyribonucleate nucleotidyltransferase; deoxynucleate polymerase; deoxyribonucleic acid duplicase; deoxyribonucleic acid polymerase; deoxyribonucleic duplicase; deoxyribonucleic polymerase; deoxyribonucleic polymerase I; DNA duplicase; DNA nucleotidyltransferase; DNA polymerase; DNA replicase; DNA-dependent DNA polymerase; duplicase; Klenow fragment; sequenase; Taq DNA polymerase; Taq Pol I; Tca DNA polymerase
Systematic Name: deoxynucleoside-triphosphate:DNA deoxynucleotidyltransferase (DNA-directed)
Purity: Verified by SDS-PAGE. Endonuclease and exonuclease activities were not detectable.
Form: Supplied as a liquid in 20mM PBS, pH 7.5, 20mM EDTA.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
1. Bollum, F.J. Calf thymus polymerase. J. Biol. Chem. 235 (1960) 2399-2403.
2. Lehman, I.R., Bessman, M.J., Simms, E.S. and Kornberg, A. Enzymatic synthesis of deoxyribonucleic acid. I. Preparation of substrates and partial purification of an enzyme from Escherichia coli. J. Biol. Chem. 233 (1958) 163-170.
3. Richardson, C.C., Schildkraut, C.L., Aposhian, H.V. and Kornberg, A. Enzymatic synthesis of deoxyribonucleic acid. XIV. Further purification and properties of deoxyribonucleic acid polymerase of Escherichia coli. J. Biol. Chem. 239 (1964) 222-232.
4. Schachman, H.K., Adler, J., Radding, C.M., Lehman, I.R. and Kornberg, A. Enzymatic synthesis of deoxyribonucleic acid. VII. Synthesis of a polymer of deoxyadenylate and deoxythymidylate. J. Biol. Chem. 235 (1960) 3242-3249.