Phusion - Revision history

Jarle Pahr: /* Thermocycling conditions */

Jarle Pahr — Wed, 13 Feb 2013 23:12:33 GMT

Thermocycling conditions

←Older revision		Revision as of 23:12, 13 February 2013
Line 52:		Line 52:
	#98C for denaturation.		#98C for denaturation.
	#Anneal at 3C above the lowest Tm if the primers are longer than 20nt, else at the Tm.		#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: [~~https~~://www.finnzymes.fi/tm_determination.html Finnzyme Tm calculator]	+	#*Note: Tm should be calculated with nearest neighbor method see: [http://www.finnzymes.fi/tm_determination.html Finnzyme Tm calculator]
	#Extend at 72C.		#Extend at 72C.

Jarle Pahr — Wed, 13 Feb 2013 16:15:49 GMT

Thermocycling conditions

←Older revision		Revision as of 16:15, 13 February 2013
Line 51:		Line 51:
	#15-30 s/kb extension time.		#15-30 s/kb extension time.
	#98C for denaturation.		#98C for denaturation.
-	#Anneal at 3C above the lowest Tm if the primers are longer ~~then~~ 20nt, ~~esle~~ at the Tm.	+	#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: [https://www.finnzymes.fi/tm_determination.html Finnzyme Tm calculator]		#*Note: Tm should be calculated with nearest neighbor method see: [https://www.finnzymes.fi/tm_determination.html Finnzyme Tm calculator]
	#Extend at 72C.		#Extend at 72C.

Jarle Pahr — Wed, 13 Feb 2013 16:01:47 GMT

Thermocycling conditions

←Older revision		Revision as of 16:01, 13 February 2013
Line 51:		Line 51:
	#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 lowest Tm if the primers are longer then 20nt, esle at the Tm.
	#*Note: Tm should be calculated with nearest neighbor method see: [https://www.finnzymes.fi/tm_determination.html Finnzyme Tm calculator]		#*Note: Tm should be calculated with nearest neighbor method see: [https://www.finnzymes.fi/tm_determination.html Finnzyme Tm calculator]
	#Extend at 72C.		#Extend at 72C.

Raik — Mon, 16 Mar 2009 14:09:22 GMT

Thermocycling conditions: comment on Tm calculation method

←Older revision		Revision as of 14:09, 16 March 2009
Line 52:		Line 52:
	#98C for denaturation.		#98C for denaturation.
	#Anneal at 3C above the Tm.		#Anneal at 3C above the Tm.
		+	#*Note: Tm should be calculated with nearest neighbor method see: [https://www.finnzymes.fi/tm_determination.html Finnzyme Tm calculator]
	#Extend at 72C.		#Extend at 72C.

Torsten Waldminghaus — Thu, 19 Feb 2009 10:36:41 GMT

Description

←Older revision		Revision as of 10:36, 19 February 2009
Line 1:		Line 1:
	==Description==		==Description==

-	Fusion of a Pyrococcus (Pfu) -like enzyme with a double-strand DNA binding domain → increased processivity.	+	Fusion of a Pyrococcus ([[Pfu]]) -like enzyme with a double-strand DNA binding domain → increased processivity.

	==Statistics about Phusion™ High-Fidelity DNA Polymerase==		==Statistics about Phusion™ High-Fidelity DNA Polymerase==

Barry Canton — Fri, 22 Feb 2008 18:07:06 GMT

←Older revision		Revision as of 18:07, 22 February 2008
Line 47:		Line 47:
	\|}		\|}

		+	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==		==Thermocycling conditions==
	#15-30 s/kb extension time.		#15-30 s/kb extension time.
Line 56:		Line 57:
	*[http://www.neb.com/nebecomm/products/productF-530.asp Purchase]		*[http://www.neb.com/nebecomm/products/productF-530.asp Purchase]
	*[http://www.neb.com/nebecomm/ManualFiles/manualF-540.pdf Manual]		*[http://www.neb.com/nebecomm/ManualFiles/manualF-540.pdf Manual]
		+	*[http://www.neb.com/nebecomm/products/productF-531.asp HF Master Mix product page]
		+	*[http://www.neb.com/nebecomm/products/productF-532.asp GC Master Mix product page]

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

Barry Canton — Thu, 19 Jul 2007 15:01:33 GMT

Thermocycling conditions

Barry Canton — Thu, 19 Jul 2007 15:00:36 GMT

Quick reference reaction mix

Barry Canton — Thu, 19 Jul 2007 14:39:42 GMT

←Older revision		Revision as of 14:39, 19 July 2007
Line 9:		Line 9:
	*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==

Reshma P. Shetty — Thu, 24 May 2007 14:22:41 GMT

←Older revision		Revision as of 14:22, 24 May 2007
Line 13:		Line 13:
	*[http://www.neb.com/nebecomm/products/productF-530.asp Purchase]		*[http://www.neb.com/nebecomm/products/productF-530.asp Purchase]
	*[http://www.neb.com/nebecomm/ManualFiles/manualF-540.pdf Manual]		*[http://www.neb.com/nebecomm/ManualFiles/manualF-540.pdf Manual]
		+
		+	[[Category:Material]] [[Category:Enzyme]] [[Category:Polymerase]] [[Category:DNA]]