Pfu DNA Polymerase


Novel enzyme for high fidelity PCR with DNA proofreading

Pfu DNA polymerase is a thermostable DNA polymerase isolated from Pyrococcus furiosus Vc1. It catalyzes the DNA-dependent polymerization of nucleotides into duplex DNA in the 5’ → 3’ direction and exhibits 3’ → 5’ exonuclease (proof reading) activity. Pfu DNA polymerase is the ideal choice for a variety of techniques requiring high-fidelity DNA synthesis by PCR reaction. It can apply to cloning, gene expression, site-directed mutagenesis and etc.

Features and Benefits
 

  • High fidelity PCR: 3’ → 5’exonuclease(proofreading) activity
  • Thermostability: Retaining 94-99% of its thermostable activity after 1 hour at 95°C.
  • Terminal Transferase Activity: Devoid of terminal transferase activity and generates blunt-ended PCR products.

 

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Enzyme Properties

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: Yes
3’ – A Overhang: No
Fragment Size: < 20 kb

 

Application

Gene synthesis
PCR or Primer extension requested High fidelity
Blunt-end PCR Cloning or mutagenesis requested high fidelity

 

Reagents Supplied

10 x Reaction Buffer: MgSO4, Tris-HCl, KCl, (NH4)2SO4,Acetylated BSA pH 8.8
dNTP Mix: 2.5 mM of each dNTP(option)

 

Concentration

2.5 U/μl



Store Temperature

-20°C


Unit Definition

One unit is defined at the amount of enzyme that will incorporate 10 nmole of dNTP into acid-insoluble material in 30 minutes at 72°C.

 

Activity Test

Amplication of limiting amounts of template DNA: comparison of Lambda DNA (A), Bacterial genomic DNA (B), Human genomic DNA (C).

* Lambda DNA was amplified using 2.5 units of enzyme in 50 uL reaction volume.
* Bactierial DNA was amplified using 2.5 units of enzyme in 50 uL reaction volume.


* Human DNA was amplified using 2.5 units of enzyme in 50 uL reaction volume.


Amplification of fragments ranging from 500 bp to 5 kb from template Lambda DNA 100 pg with 2.5 units of Pfu DNA Polymerase.
 


Notice to Purchaser

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES

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Manual

Pfu DNA Polymerase


MSDS

Pfu DNA Polymerase

Pfu DP-dilution buffer

Pfu DP-reaction buffer


Brochure

• Enzymes 2010 Brochure


Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

ISO 9001:2008 - certificate

EN ISO 13485:2003 / AC:2009 - certificate

 

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