Pfu DNA Polymerase

Name: Pfu DNA Polymerase
SKU: EMQZ1401
Availability: InStock

For research use only. Not intended for any clinical use.

Cat. No. : EMQZ1401

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Product Information

Cat. No.	EMQZ1401
Description	Pfu DNA Polymerase is ideal for amplification of DNA fragments for high-fidelity PCR applications. High fidelity is provided by a proprietary enzyme preparation containing recombinant DNA polymerase from E.coli with proofreading activity (Pyrococcus furiosis DNA Polymerase gene). Unlike Taq DNA polymerase, highly thermostable Pfu DNA polymerase possesses 3' to 5' exonuclease proofreading activity that enables the polymerase to correct nucleotide misincorporation errors. This means that Pfu DNA polymerase-generated PCR fragments will have fewer errors than Taq-generated PCR inserts. Pfu DNA Polymerase provides much higher fidelity than Taq DNA polymerase when doing PCR of problematic and / or GC-rich templates. Pfu DNA polymerase is superior for techniques that require high-fidelity DNA synthesis.
Source	An E.coli strain that carries the pfu DNA polymerase gene from Pyrococcus furiosus.
Concentration	2,500 U/ml
Applications	• High-fidelity PCR and primer-extension reactions • Generation of PCR products for cloning and expression • PCR cloning and blunt-end amplification product Generation • RT-PCR for cDNA cloning and expression • Site-directed mutagenesis • blunt-end PCR cloning
Size	100 U; 500 U
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 72°C.
Reaction Conditions	10×Pfu DNA Polymerase Reaction Buffer (containing 20 mM MgSO4): [200 mM Tris-HCl (pH 8.8 at 25°C), 100 mM (NH4)2SO4, 100 mM KCl, 1 mg/ml BSA (without nulease activity), 1% Triton X-100 (V/V), 20 mM MgSO4].
Storage	Store at -20°C

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Q & A

Customer Reviews

Customer Q&As

How many different specifications does Pfu DNA Polymerase come in, and which one is more cost-effective to buy?

A: Pfu DNA Polymerase is available in two specifications: 200U and 1000U. It is recommended to buy the 1000U specification as it provides better value with a larger package.

Does Pfu DNA Polymerase have exonuclease activity?

A: Pfu enzyme has both 3' to 5' exonuclease activity and 5' to 3' exonuclease activity but does not possess reverse transcriptase activity.

What is the fidelity of Pfu DNA Polymerase?

A: Due to the 3' to 5' exonuclease activity of Pfu enzyme, the error rate during PCR amplification is significantly reduced. The error rate of Pfu is not only much lower than Taq polymerase but also lower than some other high-fidelity DNA polymerases such as Vent, DeepVent, Pwo, Tli, etc. Therefore, Pfu enzyme is often the preferred and cost-effective choice for high-fidelity DNA amplification.

Where does Pfu DNA Polymerase originate from?

A: Pfu DNA Polymerase is expressed and purified from Escherichia coli and possesses the same properties as naturally purified Pfu DNA Polymerase.

What are the applications of Pfu DNA Polymerase?

A: Pfu DNA Polymerase can be used in high-fidelity PCR, site-directed mutagenesis, double blunt-end PCR cloning, and more.

Can Pfu DNA Polymerase be used for T-vector cloning?

A: No, Pfu enzyme cannot be used for T-vector cloning. DNA fragments amplified with Pfu enzyme have blunt ends, which are suitable for blunt-end cloning but not for traditional T-vector cloning.

Are there any specific requirements for primers when using Pfu DNA Polymerase in PCR reactions?

A: dUTP, dITP, or primers containing dUTP and dITP cannot be used in PCR amplification mediated by Pfu enzyme.

What is the purity of Pfu DNA Polymerase?

A: Pfu DNA Polymerase has good purity and does not contain DNA endonucleases, exonucleases, phosphatases, or RNases, meeting the requirements of routine PCR reactions.

How can the activity of Pfu DNA Polymerase be inhibited or deactivated?

A: Phenol-chloroform extraction can be used to inactivate Pfu enzyme.

Under what conditions is it recommended to use Pfu enzyme for reactions?

A: When amplifying DNA fragments of 2 kb or less, the amplification efficiency of Pfu enzyme is similar to Taq enzyme. However, in cases where high accuracy is required, Pfu enzyme should be chosen.

Can the buffer for Taq enzyme be used when using Pfu DNA Polymerase in PCR reactions?

A: No, you cannot use Taq enzyme's PCR buffer as a substitute for Pfu enzyme's PCR buffer. You should use the buffer provided with the Pfu enzyme product for PCR reactions.

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Customer Reviews

Good purity

Contains no DNA exonuclease, endonuclease, and phosphatase, no RNase, meeting the requirements of conventional PCR reactions.

United States

2020-08-23

Low error rate

Due to the 3' to 5' exonuclease activity of Pfu enzyme, the chance of error during the PCR amplification process is greatly reduced, with an error rate of 2.6×10-6 per nt per cycle. The error rate of Pfu not only is much lower than Taq enzyme, but also lower than some other high-fidelity DNA polymerases such as Vent, DeepVent, Pwo, Tli, etc.

United States

2021-05-12

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Pfu DNA Polymerase

Product Information

Q & A

Customer Reviews

How many different specifications does Pfu DNA Polymerase come in, and which one is more cost-effective to buy?

Does Pfu DNA Polymerase have exonuclease activity?

What is the fidelity of Pfu DNA Polymerase?

Where does Pfu DNA Polymerase originate from?

What are the applications of Pfu DNA Polymerase?

Can Pfu DNA Polymerase be used for T-vector cloning?

Are there any specific requirements for primers when using Pfu DNA Polymerase in PCR reactions?

What is the purity of Pfu DNA Polymerase?

How can the activity of Pfu DNA Polymerase be inhibited or deactivated?

Under what conditions is it recommended to use Pfu enzyme for reactions?

Can the buffer for Taq enzyme be used when using Pfu DNA Polymerase in PCR reactions?

Good purity

Low error rate

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