Paris/July 13
From 2008.igem.org
(Difference between revisions)
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* Sharing into 4 Falcon tubes of 50mL and cooling at 4°C during 40' in ice | * Sharing into 4 Falcon tubes of 50mL and cooling at 4°C during 40' in ice | ||
* Spin during 8', 2000rpm, 4°C | * Spin during 8', 2000rpm, 4°C | ||
- | * Resuspension of the cell's pellet in 20mL of cold | + | * Resuspension of the cell's pellet in 20mL of cold CaCl<sub>2</sub> (0,1M) during 30' in ice |
* Spin during 8', 2000rpm, 4°C | * Spin during 8', 2000rpm, 4°C | ||
- | * Resuspension of the cell's pellet in 1mL of cold | + | * Resuspension of the cell's pellet in 1mL of cold CaCl<sub>2</sub> (0,1M) |
* 1h of delay unexpected (problem with DNA lyophilized) | * 1h of delay unexpected (problem with DNA lyophilized) | ||
* Transformation of the Biobricks with the "classical" protocol (--> heat shock at 42°C during 40") | * Transformation of the Biobricks with the "classical" protocol (--> heat shock at 42°C during 40") |
Revision as of 19:13, 15 July 2008
delete
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
- Biobricks 2007 : use of 3µl of the minipreps
- Biobricks 2008 : use of the maximum recoverable volume of the minipreps
- 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 |