Team:Tuebingen/Notebook/Protocols/trafo
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- | <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: | + | <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a> |
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+ | <p> </p> | ||
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+ | <h3>Procedure</h3> | ||
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+ | <ol> | ||
+ | <li>Add desired volume of (plasmid) DNA (not more than 1/10 vol.) to 100 µL <i>E. coli</i> culture (i.e. to <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/chemo-competentcells">aliquot</a>) in Eppendorf-tube. </li> | ||
+ | <li>Incubate on ice for 30 min.</li> | ||
+ | <li>Heat shock at 42 °C for 90 s.</li> | ||
+ | <li>Incubate 10 min on ice.</li> | ||
+ | <li>Add 0.9 mL of <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/lb">LB-medium</a> and let bacteria grow at 37°C.</li> | ||
+ | <li>Plate on <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/plates">LB plates</a> + required amount of antibiotic and incubate at 37 °C over night.</li> | ||
+ | </ol> | ||
Revision as of 00:03, 4 October 2013
E. coli Transformation
Back to Protocols
Procedure
- Add desired volume of (plasmid) DNA (not more than 1/10 vol.) to 100 µL E. coli culture (i.e. to aliquot) in Eppendorf-tube.
- Incubate on ice for 30 min.
- Heat shock at 42 °C for 90 s.
- Incubate 10 min on ice.
- Add 0.9 mL of LB-medium and let bacteria grow at 37°C.
- Plate on LB plates + required amount of antibiotic and incubate at 37 °C over night.