Pfu DNA Polymerase

*This product has been discontinued!*

Description (Not available in USA): GenScript&39s Pfu DNA polymerase is a thermostable DNA polymerase isolated from an E. coli strain that carries the pol gene from Pyrococcus furiosus. As one of the most thermostable DNA polymerases known to have 3’ to 5’ proofreading activity, the enzyme catalyzes DNA systhesis at optimal temperature near 75°C with very low error rate; it is about eight times more accurate than Taq DNA polymerase). Pfu DNA polymerase generates blunt-ended PCR fragments and so may be useful for projects that involve blunt-end PCR cloning.

Cat. No.	E00021
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Product Introduction Applications

Description

Description (Not available in USA):

GenScript's Pfu DNA polymerase is a thermostable DNA polymerase isolated from an E. coli strain that carries the pol gene from Pyrococcus furiosus. As one of the most thermostable DNA polymerases known to have 3’ to 5’ proofreading activity, the enzyme catalyzes DNA systhesis at optimal temperature near 75°C with very low error rate; it is about eight times more accurate than Taq DNA polymerase). Pfu DNA polymerase generates blunt-ended PCR fragments and so may be useful for projects that involve blunt-end PCR cloning.

Note

Unit Definition:
one unit is defined as the amount of enzyme that can incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 74 °C.

10 X Reaction Buffer (with Mg2+):
200 mM Tris-HCl (pH 8.7 at 25°C), 100 mM (NH4)2SO4, 100 mM KCl, 1% (v/v) Triton X-100, 1 mg/ml BSA and 20 mM MgSO4

Storage Buffer and Concentration:
Supplied in 2.5 units/µl in 50 mM Tris HCl (pH 8.1), 0.1 mM EDTA, 1 mM DTT, 0.1% (v/v) Nonidet P40, 0.1% (v/v) Tween 20 and 50% (v/v) glycerol.

Product Introduction

QC Tests:

This product is tested for activity and PCR^* performance.

Figure 1. Shown below is a PCR amplification of a 544 bp gene from E. coli genomic DNA using GenScript Pfu DNA Polymerase. 0.25, 0.50, 0.75, 1.00 and 1.25 units of enzyme were used in 50 µl of PCR reaction 1 (Lane 1), 2 (Lane 2), 3 (Lane 3), 4 (Lane 4) and 5 (Lane 5), respectively. 5 µl of PCR material was loaded into each lane. M represents a 100 bp DNA marker.

Figure 2. Shown below is a comparison of GenScript Pfu to that of an anonymous competitor (A). Both enzymes are shown amplifyinga 544 bp gene from E. coli genomic DNA. 0.4, 0.6, 0.8, and 1.0 U of each enzymes was used in 50 µl of PCR material, as shown in Figure 2. 10 µl of PCR material was loaded into each lane. M represents a 100 bp DNA marker.

Storage & Stability

Storage:

Store the product at -20 °C

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Applications

Applications:

Pfu DNA Polymerase can be used in most applications including the following:

High fidelity PCR amplification.
High fidelity blunt-end cloning.
Site-directed mutagenesis.

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