Phusion
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==Thermocycling conditions== | ==Thermocycling conditions== | ||
| - | #15-30 s/kb extension time | + | #15-30 s/kb extension time. |
| - | #98C for denaturation | + | #98C for denaturation. |
#Anneal at 3C above the Tm. | #Anneal at 3C above the Tm. | ||
| + | #Extend at 72C. | ||
==References== | ==References== | ||
Revision as of 11:01, 19 July 2007
Contents |
Description
Fusion of a Pyrococcus (Pfu) -like enzyme with a double-strand DNA binding domain → increased processivity.
Statistics about Phusion™ High-Fidelity DNA Polymerase
- 10x processivity compared to Taq
- 50x fidelity compared to Taq
- Creates blunt end
- Error rate is 4.4 X 10-7 in Phusion HF buffer and 9.5 X 10-7 in GC buffer
- Non-displacing
Quick reference reaction mix
See the manual for details and special usage conditions.
| Component | Volume for 50μl reaction | Final concentration |
|---|---|---|
| 5x Phusion HF Buffer | 10μl | 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 | |
| Phusion DNA polymerase (2U/μ) | 0.5μl | 0.02 U/pl |
Thermocycling conditions
- 15-30 s/kb extension time.
- 98C for denaturation.
- Anneal at 3C above the Tm.
- Extend at 72C.
