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Pfu DNA Polymerase

Pfu DNA Polymerase

High-Fidelity PCR

  • Low error rate thermostable DNA polymerase
  • Provided with 10X buffer containing 20mM MgSO4
  • Recommended for applications that require high accuracy base insertion
  • Supplied at a concentration of 2–3u/μl

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Description

Size

Catalog number selected: M7741

Catalog number selected:  M7745

High-Fidelity PCR

 

Pfu DNA Polymerase is a high-fidelity, thermostable enzyme of approximately 90kDa isolated from Pyrococcus furiosus. The enzyme replicates DNA at 75°C, catalyzing the polymerization of nucleotides in the 5´→3´ direction in the presence of magnesium. Pfu DNA Polymerase also possesses 3´→5´ exonuclease (proofreading) activity. Base misincorporations that may occur during polymerization are rapidly excised by this proofreading activity. Consequently, Pfu DNA Polymerase is recommended for use in PCR and primer extension reactions that require high-fidelity synthesis. Pfu DNA Polymerase-generated PCR fragments are blunt-ended.

  • 10X Reaction Buffer with MgSO4: 200mM Tris-HCl (pH 8.8 at 25°C), 100mM KCl, 100mM (NH4)2SO4, 20mM MgSO4, 1.0% Triton® X-100 and 1mg/ml nuclease-free BSA.
  • Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 10nmol of dNTPs into acid-insoluble material in 30 minutes at 75°C.
  • Source: Pyrococcus furiosus strain Vc1 DSM3638.
  • Storage Buffer: 50mM Tris-HCl (pH 8.2 at 25°C), 0.1mM EDTA, 1mM DTT, 50% glycerol and 0.05% CHAPS.

Performance Guarantee: Promega PCR systems, enzymes and reagents are proven in PCR to ensure reliable, high-performance results. If you are not completely satisfied with any Promega PCR product, we will send a replacement or refund your account.

  1. Fiala, G. and Stetter, K.O. (1986) Arch. Microbiol. 145, 56.
  2. Lundberg, K.S. et al. (1991) Gene 108, 1–6.
  3. Flaman, J.M. et al. (1994) Nucl. Acids Res. 22, 3259–60.
  4. Cline, J. et al. (1996) Nucl. Acids Res. 24, 3546–51.
  5. Andre, P. et al. (1997) Genome Res. 7, 843–52.

Important Publications

With a low error rate, Pfu DNA Polymerase is optimally used for high-fidelity PCR and synthesis. Our product has been used in a wide variety of applications—such as gene cloning, gene expression or mutation analysis—across several fields of study.

Publication

Mesalam, A.A. et al. (2018) Development and characterization of a human monoclonal antibody targeting the N-terminal region of hepatitis C virus envelope glycoprotein E1. Virology 514, 30–41.

Pasello, M. et al. (2018) Construction of Human Naïve Antibody Gene Libraries. Methods in Molecular Biology 1827.

Harvey, J.A. et al. (2018) Ant-like Traits in Wingless Parasitoids Repel Attack from Wolf Spiders. Journal of Chemical Ecology 44(10), 894–904.

Dubos, M.P. et al. (2018) Characterization of a tachykinin signaling system in the bivalve mollusc Crassostrea gigas. General and Comparative Endocrinology 266, 110–118.

Tanabe, E.L.L. et al. (2018) Report of East-Central South African Chikungunya virus genotype during the 2016 outbreak in the Alagoas State, Brazil. Revista do Instituto de Medicina Tropical de São Paulo 60.

Summary

The authors used Pfu DNA Polymerase to accurately amplify and subsequently clone the envelope glycoprotein region in the hepatitis C virus (HCV) for development of a human monoclonal antibody.

The authors used Pfu DNA Polymerase to construct human naïve antibody gene libraries with high affinity and variability.

The authors used Pfu DNA Polymerase to amplify the COI gene in wingless parasitoids and subsequently clone and perform Sanger sequencing to reconstruct a partial phylogenetic tree of the organism.

The authors used Pfu DNA Polymerase to amplify the coding sequence of the Cragi-TKR gene from a cDNA library, subsequently cloned into a eukaryotic expression vector, transiently transfected using Fugene HD, and calcium responses were measured.

The authors first performed reverse transcription on Chikungunya viral RNA using ImProm-II Reverse Transcription System to generate cDNA that was then used as a template to amplify the E1 region using Pfu DNA Polymerase. The PCR amplicon was subsequently Sanger sequenced.

Protocols

Complete Protocol

Pfu DNA Polymerase Protocol

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Specifications

What’s in the box?

Item/ Catalog number selected: M7741 Part # Size
Pfu DNA Polymerase M774A 1 × 100u
Pfu 10X Reaction Buffer w/20mM MgSO4 M776A 1 × 1.2ml

SDS For  M7741

Download SDS

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

What’s in the box?

Item/ Catalog number selected: M7745 Part # Size
Pfu DNA Polymerase M774B 1 × 500u
Pfu 10X Reaction Buffer w/20mM MgSO4 M776A 3 × 1.2ml

SDS For M7745

Download SDS

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions