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Paris/July 13
From 2008.igem.org
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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
- IGEM 2008 Biobricks : use of the maximum recoverable volume minipreps
- IGEM 2007 Biobricks : use of 3µl of minipreps
| details | resitance | 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 |
