Phusion

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

A 2x supermix is now available containing either HF buffer or GC buffer, dNTPs, and Phusion polymerase. See references section below for links.

Thermocycling conditions

1. 15-30 s/kb extension time.
2. 98C for denaturation.
3. Anneal at 3C above the lowest Tm if the primers are longer then 20nt, esle at the Tm.
   • Note: Tm should be calculated with nearest neighbor method see: Finnzyme Tm calculator
4. Extend at 72C.

References

• Purchase
• Manual
• HF Master Mix product page
• GC Master Mix product page