View source for 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.''

{|{{Table}}
!Component
!Volume for 50&mu;l reaction
!Final concentration
|-
|5x Phusion HF Buffer
|10&mu;l
|1x
|-
|10mM dNTPs
|1&mu;l
|200&mu;M each
|-
|primer A
|x &mu;l
|0.5&mu;M
|-
|primer B
|x &mu;l
|0.5&mu;M
|-
|template DNA
|x&mu;l
|
|-
|H<sub>2</sub>O
|add to 50&mu;l
|
|-
|Phusion DNA polymerase (2U/&mu;)
|0.5&mu;l
|0.02 U/pl
|}

==Thermocycling conditions==
#15-30 s/kb extension time.
#98C for denaturation.
#Anneal at 3C above the Tm.
#Extend at 72C.

==References==
*[http://www.neb.com/nebecomm/products/productF-530.asp Purchase]
*[http://www.neb.com/nebecomm/ManualFiles/manualF-540.pdf Manual]

[[Category:Material]] [[Category:Enzyme]] [[Category:Polymerase]] [[Category:DNA]]