Difference between revisions of "Chemical transformation buffer comparison"

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Line 3: Line 3:
 
|'''Buffer'''
 
|'''Buffer'''
 
|'''KCl'''
 
|'''KCl'''
|'''KCH<sub>3</sub>COO'''
+
|'''KOAc'''
 
|'''CaCl<sub>2</sub>'''
 
|'''CaCl<sub>2</sub>'''
 
|'''MnCl<sub>2</sub>'''
 
|'''MnCl<sub>2</sub>'''
Line 10: Line 10:
 
|'''Glycerol'''
 
|'''Glycerol'''
 
|'''DMSO'''
 
|'''DMSO'''
|'''Sucrose'''
 
 
|'''Other'''
 
|'''Other'''
 
|'''pH'''
 
|'''pH'''
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|
 
|
 
|10%
 
|10%
|
 
 
|
 
|
 
|
 
|
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|15%
 
|15%
 
|
 
|
 +
|
 +
|pH 5.8
 +
|Hanahan patent
 +
|-
 +
|TFB-I
 +
|
 +
|30mM
 +
|20mM
 +
|50mM
 +
|100mM
 +
|
 +
|15%
 
|
 
|
 
|
 
|
 
|pH 5.8
 
|pH 5.8
|Hanahan patent
+
|
 +
|-
 +
|TFB-II
 +
|
 +
|
 +
|75mM
 +
|
 +
|10mM
 +
|
 +
|
 +
15%
 +
|
 +
|10mM MOPS
 +
|pH 6.5
 +
|
 
|-
 
|-
 
|FSB
 
|FSB
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|3mM
 
|3mM
 
|10% w/v
 
|10% w/v
|
 
 
|
 
|
 
|
 
|
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|10%
 
|10%
 
|2x 3.5%
 
|2x 3.5%
|5%
+
|5% Sucrose
|
 
 
|pH 6.4->6.2
 
|pH 6.4->6.2
 
|Jessee patent
 
|Jessee patent
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|8%
 
|8%
 
|8%
 
|8%
|
 
 
|
 
|
 
|pH 6.4->6.2
 
|pH 6.4->6.2
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|10%
 
|10%
 
|7%
 
|7%
|
 
 
|10mM MgCl<sub>2</sub>
 
|10mM MgCl<sub>2</sub>
 
|pH 6.4
 
|pH 6.4
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|
 
|
 
|15mM
 
|15mM
|15mM
+
|55mM
 
|
 
|
 
|
 
|
 
|
 
|
 
|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.
 

Latest revision as of 18:00, 2 May 2006

Buffer KCl KOAc CaCl2 MnCl2 RbCl HACoCl3 Glycerol DMSO Other pH Reference
SF 100mM 10mM 50mM 10% pH 6.1 Hanahan patent
RFT 30mM 10mM 50mM 100mM 15% pH 5.8 Hanahan patent
TFB-I 30mM 20mM 50mM 100mM 15% pH 5.8
TFB-II 75mM 10mM

15%

10mM MOPS pH 6.5
FSB 100mM 10mM 45mM 3mM 10% w/v pH 6.2 Hanahan patent
FSB 100mM 10mM 10mM 44mM ? 3mM 10% 2x 3.5% 5% Sucrose pH 6.4->6.2 Jessee patent
FSB 10mM 10mM 45mM 3mM 8% 8% pH 6.4->6.2 Stratagene '525
CCMB80 10mM 80mM 20mM 10% 7% 10mM MgCl2 pH 6.4 Invitrogen '464
TB 250mM 15mM 55mM 7% 10mM PIPES pH 6.7 pH 5.7->6.2 Inoue90