Pfu Polymerases

Pfu Polymerase
Pfu DNA polymerase has superior thermstability and proofreading properties compared to other thermostable polymerases. Pfu DNA polymerase-generated PCR fragments will have fewer errors than Taq-generated PCR inserts. Using Pfu DNA polymerase in your PCR reactions results in blunt-ended PCR products, which are ideal for cloning into blunt-ended vectors. Pfu DNA polymerase is superior for techniques that require high-fidelity DNA synthesis.

High-fidelity PCR and primer-extension reactions
High fidelity PCR for cloning into blunt-ended vectors
Site-directed mutagenesis

Pfu Polymerase
Source                                    Pyrococcus furiosus
Molecular Weight             90 kDa

Elongation Velocity          0.2 –  0.4 kb/min (70-75°C)
Amplification Capacity   Up to 5 kb (simple template)
Polymerase activity          5′ to 3′ polymerase activity
Exonuclease activity         Lacks 3′-5′ exonuclease proof reading 

High fidelity: 10x more powerful than regular Taq Polymerase
Thermostable: 94% activity after 2 hour incubation at 95˚C vectors
Blunt-end PCR: for direct PCR cloning
Proof reading: corrects for nucleotide-mis incorporation errors

Storage Buffer
20mM Tris HCl (pH 8.0)
100 mM KCl
3 mM MgCl2
1 mM DTT
0.1% NP-40
0.1% Tween 20
0.2 mg/ml BSA
50% (v/v) glycerol

10X PCR Buffer with Mg2+
200 mM Tris-HCl (pH 8.8)
100 mM KCl
100 mM (NH4)2SO4
1% Triton X-100
1mg/ml BSA

Catalog # Size Price
TP-25-01 500 U $ 60.00
TP-25-02 1000 U $ 110.00

Pfu Polymerase Mix
Pfu Mix (2X) is a pre-mixed, ready-to-use solution containing Pfu DNA Polymerase, dNTPs, MgSO4 and Reaction Buffer at optimal concentrations for efficient amplification of DNA templates by PCR. To prepare the final PCR, primers and template DNA are added to the aliquoted Pfu Mix. Pfu Mix contributes to highly reproducible PCR by reducing the risk of pipetting errors, miscalculation and contamination. It also contributes to higher sensitivity by adding intensifier and optimizer.

High-fidelity PCR and primer-extension reactions
Routine PCR with high reproducibility
Generation of PCR products for TA cloning
Site-directed mutagenesis

Composition of 2X Pfu Mix
0.15U/μl Pfu DNA Polymerase
2x Pfu Buffer
0.4 mM dNTPs
4 mM MgSO4
0.02% Bromophenol blue

Pfu DNA Polymerase in a ready-to-use mix
Higher yields with minimal optimization
Reduced hands-on time
Lower contamination risks
Reduced pipetting errors

Catalog # Size Price
TP-26-01 1 ml $78.00
TP-26-02 1 x 5 ml $ 351.00

Unit Definition
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmole of dNTPs inot an acid-insoluble form in 30 minutes at 70˚C using hering sperm DNA as substrate.

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