Pfu DNA polymerase, derived from the hyperthermophilic archae Pyrococcus furiosus, has been shown to exhibit superior thermostability and proofreading properties compared to other thermostable polymerase. 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. 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.
Components: Pfu DNA polymerase (2.5 U/ul or 5 U/ul-please see user manual), 10x Pfu buffer
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
PCR cloning and blunt-ended amplification product generation
Unit Definition
One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nM of dNTPs into an acid-insoluble form in 30 minutes at 70°C using hering sperm DNA as substrate.
Store all components at –20°C
