Phusion: Difference between revisions
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*Error rate is 4.4 X 10-7 in Phusion HF buffer and 9.5 X 10-7 in GC buffer | *Error rate is 4.4 X 10-7 in Phusion HF buffer and 9.5 X 10-7 in GC buffer | ||
*Non-displacing | *Non-displacing | ||
==Quick reference reaction mix== | |||
''See the manual for details and special usage conditions.'' | |||
{|{{Table}} | |||
!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 | |||
| | |||
|- | |||
|H<sub>2</sub>O | |||
|add to 50μl | |||
| | |||
|- | |||
|Phusion DNA polymerase (2U/μ) | |||
|0.5μl | |||
|0.02 U/pl | |||
|} | |||
==References== | ==References== | ||
Revision as of 07:39, 19 July 2007
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 |
