In the experiment, the 773bp target fragment was amplified by PCR using PUC57 as the template. At the same time, competing enzymes were added to investigate the enzyme activity of Pfu DNA Polymerase. As shown in the figure, this product has the following effects.
Lane 1 Negative Control (negative control with no added enzyme only);
Lane 2 UA-Pfu DNA Polymerase 2.5U;
Lane 3 UA-Pfu DNA Polymerase 5U;
Lane 4 Competing product A 5U;
Lane 5 Competing product B 5U;
Pfu DNA Polymerase
Pfu DNA Polymerase
Price:
Regular price
$164 USD
Regular price
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$164 USD
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Product Details
Product Details
Product Specification
| Species | Pyrococcus furiosus |
| Synonyms | DNA polymerase、 DNA polymerase B、Pfu polymerase、Pol I |
| Expression System | E.coli |
| Molecular Weight | 90 kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Tag | His Tag |
| Storage Buffer | 20 mM Tris-HCl, 0.1 mM EDTA, 0.1% Tween20, 0.1% triton X100, 1 mM DTT, 100 mM KCl, 50%Glycerol、pH 8.2 @ 25°C |
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference | [1] Picard Véronique, et al. "A rapid and efficient one-tube PCR-based mutagenesis technique using Pfu DNA polymerase." Nucleic Acids Research 22.13(1994):2587-91. |
Background
Pfu DNA Polymerase is a thermostable enzyme that replicates DNA at 75°C. It catalyzes the polymerization of nucleotides into duplex DNA in the 5´→3´ direction in the presence of magnesium. The enzyme has a molecular weight of approximately 90,000 daltons as estimated from the predicted amino acid sequence and exhibits 3´→5´ exonuclease (proofreading) activity. Pfu DNA Polymerase is recommended for use in PCR and primer extension reactions that require high fidelity.
Components
Storage Solution : 5 U/ul Pfu DNA Polymerase, 20 mM Tris-HCl, 0.1 mM EDTA, 0.1% Tween20, 0.1% triton X100, 1 mM DTT, 100 mM KCl, 50%Glycerol、pH 8.2 @ 25°C
10* Reaction Buffer: 200 mM Tris-HCl (pH 8.8) , 20 mM MgSO4, 100 mM KCl, 100 mM (NH4) 2SO4, 1% Triton X-100, 1 mg/ml nuclease-free BSA
Guidelines
Note: 1. It is critical to withhold Pfu DNA Polymerase until after the addition of dNTPs; otherwise, the proofreading activity of the polymerase may degrade the primers, resulting in nonspecific amplification and reduced product yield.
2. Assemble on ice.
Unit Definition
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 74°C
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Bioactivity
SDS-PAGE
1μg (R: reducing condition, N: non-reducing condition).
SEC-HPLC
