Team:EPF-Lausanne/Protocols/Transformation
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- | + | :1. Thaw cells on ice for 20min | |
- | + | :2. Add DNA (ligation product) to the cells, mix. | |
- | + | :3. Incubate 20min on ice | |
- | + | :4. Heat shock 45s @ 42°C | |
- | + | :5. Incubate 20min on ice (optional) | |
- | + | :6. Add 500 µl SOC and incubate 1h @ 37°C (shaking) | |
- | + | :7. Spread over LB-agar plate | |
- | + | :8. Incubate overnight @ 37°C | |
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Latest revision as of 18:46, 20 October 2009
Material
- 50 µl competent cells
- LB-agar plates with corresponding antibiotics
- Water bath at 42°C
Procedure
- 1. Thaw cells on ice for 20min
- 2. Add DNA (ligation product) to the cells, mix.
- 3. Incubate 20min on ice
- 4. Heat shock 45s @ 42°C
- 5. Incubate 20min on ice (optional)
- 6. Add 500 µl SOC and incubate 1h @ 37°C (shaking)
- 7. Spread over LB-agar plate
- 8. Incubate overnight @ 37°C