Pfu DNA polymerase is a thermostable enzyme isolated from Pyrococcus furiousus. The enzyme replicates DNA at 75°C, catalyzing the polymerization of nucleotides into duplex DNA in the 5'→3' direction.Pfu DNA polymerase possesses 3'→5' exonuclease (proofreading) activity. Base misincorporation is rapidly excised by the proofreading activity of the polymerase. Pfu DNA polymerase is recommended for PCR and primer extension reactions that require high- fidelity. The fragments of Pfu DNA polymerase generated are blunt-ended.
Error rate: 2 x10-6
Unit description
One unit of Pfu DNA Polymerase incorporates 10 nmol of dNTP into acid-insoluble material in 30 min at 74°C.
Storage buffer:
50% glycerol (v/v), 20 mM Tris-HCl pH 8.7 at -20ºC, 100 mM KCl, 0.1 mM EDTA.
Source: E coli clone
PCR result (Figure 1.)
Pfu DNA polymerase amplified 1kb to 7.5kb DNA fragement.
PCR cycles program (Table 1.)
IMPORTANT: Annealing temperature should be 2-6ºC lower than the primer melting temperature.
Shipping and Storage conditions
Shipping and temporary storage at -20°C and for up to 1 month at room temperature has no detrimental effects on the quality of Pfu DNA polymerase.
Quality control
The enzyme is free of nicking and priming activities, exonucleases and non-specific endonucleases. SDS/PAGE - 95 kD band. Activity and stability tested via thermo-cycling.