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Team:INSA-Lyon/Protocols/Transformation
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<h3> Protocols </h3> | <h3> Protocols </h3> | ||
| - | + | <br><p>Choose a protocol to read its description : | |
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<h5 style="text-indent:20px">Transformation</h5> | <h5 style="text-indent:20px">Transformation</h5> | ||
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Always keep competent bacteria on ice before the heat shock in order to maintain competence. | Always keep competent bacteria on ice before the heat shock in order to maintain competence. | ||
<li>Add 1 µL of DNA (can be adjust with the DNA concentration, for example transform the entire ligation product) to 200 µl of competent cells. Mix carefully by moving the tip around.</li> | <li>Add 1 µL of DNA (can be adjust with the DNA concentration, for example transform the entire ligation product) to 200 µl of competent cells. Mix carefully by moving the tip around.</li> | ||
Latest revision as of 21:03, 25 October 2010
Protocols
Choose a protocol to read its description :
- Competent cells
- Transformation
- DNA extraction
- Digestion
- Ligation
- Measure of temperature and shaking speed influence
- Measure of osmotic pressure influence
- Granules extraction and intein cleavage
- Biofilms quantification
- Extra
Transformation
-
Always keep competent bacteria on ice before the heat shock in order to maintain competence.
- Add 1 µL of DNA (can be adjust with the DNA concentration, for example transform the entire ligation product) to 200 µl of competent cells. Mix carefully by moving the tip around.
- Incubate cells on ice for 30 min.
- Heat-shock cells for 2 min in a 42°C water bath.
- Incubate cells on ice for 5 min.
- Add 0.8 ml of room temperature LB.
- Incubate 1h at 37°C.
- Spread 100 µl of bacterial suspension on a LB agar plate containing the appropriate antibiotic and the rest in an other plate LB+AB
- Incubate overnight at 37°C
