Paris/July 13

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Revision as of 18:35, 15 July 2008

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Preparation of Competent Cells

Dilution 1/200e (1mL of the O/N culture on 200mL of LB)
Culture at 37°C until DO=0,57 (around 3h)
Sharing into 4 Falcon tubes of 50mL and cooling at 4°C during 40' in ice
Spin during 8', 2000rpm, 4°C
Resuspension of the cell's pellet in 20mL of cold CaCl2 (0,1M) during 30' in ice
Spin during 8', 2000rpm, 4°C
Resuspension of the cell's pellet in 1mL of cold CaCl2 (0,1M)
1h of delay unexpected (problem with DNA lyophilized)
Transformation of the Biobricks with the "classical" protocol (--> heat shock at 42°C durin 40")
Incubation O/N at 37°C of the transformed bacteria

Remark :

Biobricks 2008 : use of the maximum recoverable volume minipreps
Biobricks 2007 : use of 3µl of minipreps

Liste of the transformation:

Biobricks 2007

Strain	Transformation
MP1 (= J23100)	ok
MP2 (= J23107)	ok
MP3 (= B0015)	ok
MP5 (= JB0030)	ok
MP6 (= E0422)	ok

Biobricks 2008

details	Strain	Antibio	Plate 2007
2E_SP 1000	B0034	A	3O P1
10F_SP 1001	R0079	A	19N P1
4F_SP 1013	C0079	K	7K P2
6H_SP 1001	C0179	A	6H P1
6D_SP 1007	S03879	A	/
6B_SP 1001	S03154	A	24K P1
10F_SP 1002	J23101	A	21G P3
11F_SP 1002	J23109	A	23G P3
4C_SP 1000	R0040	A	7O P1
2G_SP 1000	C0040	A	5C P1
1C_SP 1008	PSB1A3	A	23E P1
7G_SP 1013	E1010	K	15M P2
7E_SP 1001	E0040	A	5H P1
2F_SP 1016	E0030	K	7G P1

@@ Line 24: / Line 24: @@
 [[Liste of the transformation:]]
 '''Biobricks 2007'''
 {| border="1"
@@ Line 49: / Line 50: @@
 |align="center"|details
 |align="center"|Strain
-|align="center"|Antibiotics
+|align="center"|Antibio
 |align="center"|Plate 2007
 |-
-|2E_SP 1000
+|align="center"|2E_SP 1000
-|B0034
+|align="center"|B0034
-|A
+|align="center"|A
-|3O P1
+|align="center"|3O P1
 |-
-|10F_SP 1001
+|align="center"|10F_SP 1001
-|R0079
+|align="center"|R0079
-|A
+|align="center"|A
-|19N P1
+|align="center"|19N P1
 |-
 |4F_SP 1013
-|C0079
+|align="center"|C0079
-|K
+|align="center"|K
-|7K P2
+|align="center"|7K P2
 |-
-|6H_SP 1001
+|align="center"|6H_SP 1001
-|C0179
+|align="center"|C0179
-|A
+|align="center"|A
-|6H P1
+|align="center"|6H P1
 |-
 |6D_SP 1007
-|S03879
+|align="center"|S03879
-|A
+|align="center"|A
-| /
+|align="center"| /
 |-
-|6B_SP 1001
+|align="center"|6B_SP 1001
-|S03154
+|align="center"|S03154
-|A
+|align="center"|A
-|24K P1
+|align="center"|24K P1
 |-
-|10F_SP 1002
+|align="center"|10F_SP 1002
-|J23101
+|align="center"|J23101
-|A
+|align="center"|A
-|21G P3
+|align="center"|21G P3
 |-
-|11F_SP 1002
+|align="center"|11F_SP 1002
-|J23109
+|align="center"|J23109
-|A
+|align="center"|A
-|23G P3
+|align="center"|23G P3
 |-
-|4C_SP 1000
+|align="center"|4C_SP 1000
-|R0040
+|align="center"|R0040
-|A
+|align="center"|A
-|7O P1
+|align="center"|7O P1
 |-
-|2G_SP 1000
+|align="center"|2G_SP 1000
-|C0040
+|align="center"|C0040
-|A
+|align="center"|A
-|5C P1
+|align="center"|5C P1
 |-
-|1C_SP 1008
+|align="center"|1C_SP 1008
-|PSB1A3
+|align="center"|PSB1A3
-|A
+|align="center"|A
-|23E P1
+|align="center"|23E P1
 |-
 |7G_SP 1013
-|E1010
+|align="center"|E1010
-|K
+|align="center"|K
-|15M P2
+|align="center"|15M P2
 |-
 |7E_SP 1001
-|E0040
+|align="center"|E0040
-|A
+|align="center"|A
-|5H P1
+|align="center"|5H P1
 |-
 |2F_SP 1016
-|E0030
+|align="center"|E0030
-|K
+|align="center"|K
-|7G P1
+|align="center"|7G P1
 |}