From 2008.igem.org
minipreps
souche stockage
MiniPreps
- Use of Promega's protocol on all the clones cultivated on the 24th.
- Preparation of 50µl of minipreps in simple.
List of Minipreps
| Name
| Biobrick
| Description
|
| MP100
| [http://partsregistry.org/Part:BBa_B0034 B0034]
| Strongest RBS (Efficiency = 1)
|
Transformations
Protocol
use of TOP10 Chemically competent cells
- Defroze competent cells on ice during 5'
- Add 5µl of DNA Ligation in 50µL of competent bacterias (or 1µL for the positive control puc19)
- Incubate 30' on ice
- Heat-shock the cells during 30" at 42°C without shaking
- Put 2' on ice
- Add 250µL of pre-warmed SOC medium (4°C)
- Incubate 1h at 37°C under shaking (225rpm)
- Spin at 5.000rpm during 30"
- Remove 150µL of supernatant
- Resuspent the pellet in the 150µL left
- Spread on adequated plates
- Incubate O/N at 37°C
List of transformation
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