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
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|
|H2O||add to 50μl|
|Phusion DNA polymerase (2U/μ)||0.5μl||0.02 U/pl|
A 2x supermix is now available containing either HF buffer or GC buffer, dNTPs, and Phusion polymerase. See references section below for links.
- 15-30 s/kb extension time.
- 98C for denaturation.
- Anneal at 3C above the lowest Tm if the primers are longer than 20nt, else at the Tm.
- Note: Tm should be calculated with nearest neighbor method see: Finnzyme Tm calculator
- Extend at 72C.