Description |
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 mins 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
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.
Cat No |
Pack size |
Conc. |
TAMB04Z-500 | 500U | 5U/uL |
TAMB04Z-2500 | 2500U | 5U/uL |
PCR cycles program |
Step | Temperature | Time | Cycle |
Initial denaturation | 94-95ºC | 1-3 mins | 1 |
Denaturation | 94-95ºC | 0.2-1 | 25-35 |
Annealing | 50-68ºC | 0.2-1 | |
Extension | 68-75ºC | 2mins/1kb | |
Final extension | 68-75ºC | 5 | 1 |
IMPORTANT: Annealing temperature should be 2-6ºC lower than the primer melting temperature.
Shipping and Storage conditions |
Shipping and temporary storage at -20 and for up to 1 month at room temperature has no detrimental effects on the quality of Pfu DNA polymerase.