Pfx50
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Description
Fusion of a Thermococcus zilligii polymerase with an accessory protein. Contains proofreading 3'->5' exonuclease activity. Accesory protein stabilizes primer-template complexes in PCR
Statistics about Pfx50 Polymerase
- 50x fidelity compared to Taq
- 15 second per kb, ideal 60 sec per kb
Quick reference reaction mix
See the manual for details and special usage conditions.
| Component | Volume for 50μl reaction | Final concentration |
|---|---|---|
| 10x Pfx50 PCR mix | 5μl | 1x |
| 10mM dNTPs | 1μl | 200μM each |
| primer A | x μl | 0.5μM |
| primer B | x μl | 0.5μM |
| template DNA | xμl | |
| H2O | add to 50μl | |
| Pfx50 DNA polymerase (5U/μ) | 0.5μl | 0.02 U/pl |
Thermocycling conditions
- 15-30 s/kb extension time.
- 98C for denaturation.
- Anneal at 2C lower than the Tm.
- Note: Tm should be calculated with nearest neighbor method see: Finnzyme Tm calculator
- Extend at 72C.
