Chemical transformation buffer comparison - Revision history

Tk at 01:00, 3 May 2006

Tk — Wed, 03 May 2006 01:00:17 GMT

Tk — Tue, 02 May 2006 20:09:10 GMT

Tk — Tue, 02 May 2006 19:17:01 GMT

Tk — Tue, 02 May 2006 18:46:26 GMT

←Older revision		Revision as of 18:46, 2 May 2006
Line 104:		Line 104:
	\|		\|
	\|15mM		\|15mM
-	\|~~15mM~~	+	\|55mM
	\|		\|
	\|		\|
Line 110:		Line 110:
	\|7%		\|7%
	\|		\|
-	\|	+	\|10mM PIPES pH 6.7
	\|pH 5.7->6.2		\|pH 5.7->6.2
	\|Inoue90		\|Inoue90
	\|}		\|}
-
-
-	Hanahan suggests that DH5a and derivatives are best prepared chemically with the TSB or FSB buffer protocols, while the MC1061 derivative strains such as DH10B and Top10 are best prepared using CCMB80 protocols. Also, the MC1061 strains should not be grown in Mg<sup>++</sup> containing culture media in preparation for transformation.

Tk — Tue, 02 May 2006 18:25:04 GMT

←Older revision		Revision as of 18:25, 2 May 2006
Line 96:		Line 96:
	\|7%		\|7%
	\|		\|
-	\|	+	\|10mM MgCl<sub>2</sub>
	\|pH 6.4		\|pH 6.4
	\|Invitrogen '464		\|Invitrogen '464
Line 114:		Line 114:
	\|Inoue90		\|Inoue90
	\|}		\|}
		+
		+
		+	Hanahan suggests that DH5a and derivatives are best prepared chemically with the TSB or FSB buffer protocols, while the MC1061 derivative strains such as DH10B and Top10 are best prepared using CCMB80 protocols. Also, the MC1061 strains should not be grown in Mg<sup>++</sup> containing culture media in preparation for transformation.

Tk — Tue, 02 May 2006 17:33:31 GMT

Tk — Tue, 02 May 2006 17:22:23 GMT

←Older revision		Revision as of 17:22, 2 May 2006
Line 1:		Line 1:
-	Buffer KCl ~~KCH3COO CaCl2 MnCl2~~ RbCl Glycerol ~~HACoCl3~~ DMSO Sucrose pH ~~Ref~~	+	{\| border="1"
-		+	\|-
-	SF ~~100uM 10uM 50uM~~ 10% 6.1 Hanahan patent	+	\|'''Buffer'''
-	RFT ~~30uM 10uM 50uM 100uM~~ 15% 5.8 Hanahan patent	+	\|'''KCl'''
-	FSB 100mM 10mM 45mM 10% w/v ~~3mM yes~~ 6.2 Hanahan patent	+	\|'''KCH<sub>3</sub>COO'''
		+	\|'''CaCl<sub>2</sub>'''
		+	\|'''MnCl<sub>2</sub>'''
		+	\|'''RbCl'''
		+	\|'''HACoCl<sub>3</sub>'''
		+	\|'''Glycerol'''
		+	\|'''DMSO'''
		+	\|'''Sucrose'''
		+	\|'''Other'''
		+	\|'''pH'''
		+	\|'''Reference'''
		+	\|-
		+	\|SF
		+	\|100mM
		+	\| 10mM
		+	\| 50mM
		+	\|
		+	\|
		+	\|
		+	\|10%
		+	\|
		+	\|
		+	\|
		+	\|pH 6.1
		+	\|Hanahan patent
		+	\|-
		+	\|RFT
		+	\|
		+	\| 30mM
		+	\| 10mM
		+	\|50mM
		+	\|100mM
		+	\|
		+	\|15%
		+	\|
		+	\|
		+	\|
		+	\|pH 5.8
		+	\|Hanahan patent
		+	\|-
		+	\|FSB
		+	\|100mM
		+	\|10mM
		+	\|
		+	\|45mM
		+	\|
		+	\|3mM
		+	\|10% w/v
		+	\|
		+	\|
		+	\|
		+	\|pH 6.2
		+	\|Hanahan patent
		+	\|-
		+	\|FSB
		+	\|100mM
		+	\|10mM
		+	\|10mM
		+	\|44mM
		+	\|?
		+	\|3mM
		+	\|10%
		+	\|2x 3.5%
		+	\|5%
		+	\|
		+	\|pH 6.4->6.2
		+	\|Jessee patent

-	FSB 100mM 10mM 10mM ~~44mM ? 10%~~ 3mM 2x 3.5% 5% 6.4 -> 6.2 ~~Jesse patent~~	+	\|-
-	~~FSB 100mM 10mM 10mM 45mM 8% 3mM 2x 3.5% 5% 6.4~~ Invitrogen '525	+	\|FSB
-	~~(add 50 uM b~~-~~ME and 110mM NaCl, incubate on ice for 10 min, add DNA, 1/2 hr on ice, heat shock)~~	+	\|100mM
-	CCMB80 10mM 80mM 20mM 10% 6.4 Invitrogen '464 ~~(10 mM MgCl2)~~	+	\|10mM
-	TB 250mM 15mM 15mM 7% 6.7 -> 6.2 ~~Inoue~~	+	\|10mM
		+	\|45mM
		+	\|
		+	\|3mM
		+	\|8%
		+	\|2x 3.5%
		+	\|5%
		+	\|
		+	\|pH 6.4->6.2
		+	\|Invitrogen '525
		+	\|-
		+	\|CCMB80
		+	\|
		+	\|10mM
		+	\|80mM
		+	\|20mM
		+	\|
		+	\|
		+	\|10%
		+	\|7%
		+	\|
		+	\|
		+	\|pH 6.4
		+	\|Invitrogen '464
		+	\|-
		+	\|TB
		+	\|250mM
		+	\|
		+	\|15mM
		+	\|15mM
		+	\|
		+	\|
		+	\|
		+	\|7%
		+	\|
		+	\|
		+	\|pH 5.7->6.2
		+	\|Inoue90
		+	\|}

Tk — Tue, 02 May 2006 00:33:25 GMT

←Older revision		Revision as of 00:33, 2 May 2006
Line 9:		Line 9:
	(add 50 uM b-ME and 110mM NaCl, incubate on ice for 10 min, add DNA, 1/2 hr on ice, heat shock)		(add 50 uM b-ME and 110mM NaCl, incubate on ice for 10 min, add DNA, 1/2 hr on ice, heat shock)
	CCMB80 10mM 80mM 20mM 10% 6.4 Invitrogen '464 (10 mM MgCl2)		CCMB80 10mM 80mM 20mM 10% 6.4 Invitrogen '464 (10 mM MgCl2)
		+	TB 250mM 15mM 15mM 7% 6.7 -> 6.2 Inoue

Tk — Mon, 01 May 2006 03:35:30 GMT

←Older revision		Revision as of 03:35, 1 May 2006
Line 8:		Line 8:
	FSB 100mM 10mM 10mM 45mM 8% 3mM 2x 3.5% 5% 6.4 Invitrogen '525		FSB 100mM 10mM 10mM 45mM 8% 3mM 2x 3.5% 5% 6.4 Invitrogen '525
	(add 50 uM b-ME and 110mM NaCl, incubate on ice for 10 min, add DNA, 1/2 hr on ice, heat shock)		(add 50 uM b-ME and 110mM NaCl, incubate on ice for 10 min, add DNA, 1/2 hr on ice, heat shock)
		+	CCMB80 10mM 80mM 20mM 10% 6.4 Invitrogen '464 (10 mM MgCl2)

Tk — Sun, 30 Apr 2006 23:03:33 GMT

←Older revision		Revision as of 23:03, 30 April 2006
Line 5:		Line 5:
	FSB 100mM 10mM 45mM 10% w/v 3mM yes 6.2 Hanahan patent		FSB 100mM 10mM 45mM 10% w/v 3mM yes 6.2 Hanahan patent

-	FSB 100mM 10mM 10mM 44mM ? 10% 3mM 3.~~5x2~~% 5% 6.4 -> 6.2 Jesse patent	+	FSB 100mM 10mM 10mM 44mM ? 10% 3mM 2x 3.5% 5% 6.4 -> 6.2 Jesse patent
		+	FSB 100mM 10mM 10mM 45mM 8% 3mM 2x 3.5% 5% 6.4 Invitrogen '525
		+	(add 50 uM b-ME and 110mM NaCl, incubate on ice for 10 min, add DNA, 1/2 hr on ice, heat shock)