Paris/July 13

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Revision as of 08:22, 16 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 during 40")
Incubation O/N at 37°C of the transformed bacteria

Remark :

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

Liste of the transformation:

Biobricks 2007

miniprep	details	Transformation
MP1	J23100	ok
MP2	J23107	ok
MP3	B0015	ok
MP5	JB0030	ok
MP6	E0422	ok

Biobricks 2008

book	plasmid	Antibio	Plate 2007	Results
2E_SP 1000	B0034	A	3O P1	No transformation
10F_SP 1001	R0079	A	19N P1	No transformation
4F_SP 1013	C0079	K	7K P2	No transformation
6H_SP 1001	C0179	A	6H P1	No transformation
6D_SP 1007	S03879	A	/	No transformation
6B_SP 1001	S03154	A	24K P1	No transformation
10F_SP 1002	J23101	A	21G P3	No transformation
11F_SP 1002	J23109	A	23G P3	No transformation
4C_SP 1000	R0040	A	7O P1	No transformation
2G_SP 1000	C0040	A	5C P1	No transformation
1C_SP 1008	PSB1A3	A	23E P1	No transformation
7G_SP 1013	E1010	K	15M P2	No transformation
7E_SP 1001	E0040	A	5H P1	No transformation
2F_SP 1016	E0030	K	7G P1	No transformation

@@ Line 28: / Line 28: @@
 *'''Biobricks 2007'''
 {| border="1"
-|align="center"|Strain
+|align="center"|miniprep
+|align="center"|details
 |align="center"|Transformation
 |-
-|MP1 (= J23100)
+|align="center"|MP1
+|align="center"|J23100
 |align="center"|ok
 |-
-|MP2 (= J23107)
+|align="center"|MP2
+|align="center"|J23107
 |align="center"|ok
 |-
-|MP3 (= B0015)
+|align="center"|MP3
+|align="center"|B0015
 |align="center"|ok
 |-
-|MP5 (= JB0030)
+|align="center"|MP5
+|align="center"|JB0030
 |align="center"|ok
 |-
-|MP6 (= E0422)
+|align="center"|MP6
+|align="center"|E0422
 |align="center"|ok
 |}
@@ Line 49: / Line 55: @@
 *'''Biobricks 2008'''
 {| border="1"
-|align="center"|details
+|align="center"|book
-|align="center"|Strain
+|align="center"|plasmid
 |align="center"|Antibio
 |align="center"|Plate 2007
+|align="center"|Results
 |-
 |align="center"|2E_SP 1000
@@ Line 58: / Line 65: @@
 |align="center"|A
 |align="center"|3O P1
+|align="center"|No transformation
 |-
 |align="center"|10F_SP 1001
@@ Line 63: / Line 71: @@
 |align="center"|A
 |align="center"|19N P1
+|align="center"|No transformation
 |-
 |align="center"|4F_SP 1013
@@ Line 68: / Line 77: @@
 |align="center"|K
 |align="center"|7K P2
+|align="center"|No transformation
 |-
 |align="center"|6H_SP 1001
@@ Line 73: / Line 83: @@
 |align="center"|A
 |align="center"|6H P1
+|align="center"|No transformation
 |-
 |align="center"|6D_SP 1007
@@ Line 78: / Line 89: @@
 |align="center"|A
 |align="center"| /
+|align="center"|No transformation
 |-
 |align="center"|6B_SP 1001
@@ Line 83: / Line 95: @@
 |align="center"|A
 |align="center"|24K P1
+|align="center"|No transformation
 |-
 |align="center"|10F_SP 1002
@@ Line 88: / Line 101: @@
 |align="center"|A
 |align="center"|21G P3
+|align="center"|No transformation
 |-
 |align="center"|11F_SP 1002
@@ Line 93: / Line 107: @@
 |align="center"|A
 |align="center"|23G P3
+|align="center"|No transformation
 |-
 |align="center"|4C_SP 1000
@@ Line 98: / Line 113: @@
 |align="center"|A
 |align="center"|7O P1
+|align="center"|No transformation
 |-
 |align="center"|2G_SP 1000
@@ Line 103: / Line 119: @@
 |align="center"|A
 |align="center"|5C P1
+|align="center"|No transformation
 |-
 |align="center"|1C_SP 1008
@@ Line 108: / Line 125: @@
 |align="center"|A
 |align="center"|23E P1
+|align="center"|No transformation
 |-
 |align="center"|7G_SP 1013
@@ Line 113: / Line 131: @@
 |align="center"|K
 |align="center"|15M P2
+|align="center"|No transformation
 |-
 |align="center"|7E_SP 1001
@@ Line 118: / Line 137: @@
 |align="center"|A
 |align="center"|5H P1
+|align="center"|No transformation
 |-
 |align="center"|2F_SP 1016
@@ Line 123: / Line 143: @@
 |align="center"|K
 |align="center"|7G P1
+|align="center"|No transformation
 |}