DNA Safe Stains
Pfu DNA polymerase
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 nmolof 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
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 | 94-95°C | 1-3 mins | 1 |
| Denaturation | 94-95°C | 0.2-1 | 25-35 |
| Annealing | 50-68°C | 0.2-1 | |
| Extension | 68-75°C | 2min/1kb | |
| Final extension | 68-75°C | 5 | 1 |
| IIMPORTANT: 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.
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