Pfu DNA polymerase, derived from the hyperthermophilic archae Pyrococcus furiosus, has superior thermostability and proofreading properties compared to other thermostable polymerase. Its molecular weight is 90 kD. It can amplify DNA target up to 2kb. The elongation velocity is 0.2~0.4kb/min (70~75°C). 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. 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.
STORAGE BUFFER
20mM Tris.HCl (pH8.0), 100mM KCl, 3mM MgCl2, 1mM DTT, 0.1% Nonidet® P-40, 0.1% Tween® 20, 0.2mg/ml BSA, 50% (v/v) glycerol.
10X Pfu BUFFER
200mM Tris-HCl (pH8.8), 100mM KCl, 100mM (NH4)2SO4, 20mM MgSO4, 1% Triton® X-100 and 1mg/ml BSA.
UNIT DEFINITION
One unit (U) of enzyme is defined as the amount of enzyme needed to catalyze the incorporation of 10 nanomoles of deoxyribonucleotides into acid-insoluble material in 30 minutes at 70°C using herring sperm DNA as a substrate.
A 2X mastermix Pfu polymerase is also available.
Taq polymerase is also available
Features
- 3'-5' exonuclease activity provides a low error rate
- One of the most thermostable DNA polymerases known
- Lack of extendase activity means no unwanted 3’ overhangs
- Optimal for blunt-ended PCR cloning
- Optimum temperature near 75°C
- 95% active after 1hour incubation at 98°C
Applications
- High-fidelity PCR and primer-extension reactions
- High fidelity PCR for cloning into blunt-ended vectors.
- Site-directed mutagenesis.
