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Team:Grenoble/Biology/Protocols/Transformation 2
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<h1>Transformation of <i>E. coli</i> competent cells (heat shock)</h1> | <h1>Transformation of <i>E. coli</i> competent cells (heat shock)</h1> | ||
<h2>Goal</h2> | <h2>Goal</h2> | ||
| - | Transform <i>E. coli</i> competent cells | + | Transform <i>E. coli</i> competent cells with foreign DNA. |
</section> | </section> | ||
<br/> | <br/> | ||
Revision as of 12:48, 27 August 2012
Transformation of E. coli competent cells (heat shock)
Goal
Transform E. coli competent cells with foreign DNA.Protocol
Following are the instructions from the OpenWetWare website.- Thaw TSS cells on ice.
- Add DNA, pipette gently to mix (1 µL of prepped plasmid is more than enough).
- Let sit for 30 minutes on ice.
- Incubate cells for 30 seconds at 42°C.
- Incubate cells on ice for 2 min.
- Add 1 mL of LB medium at room temperature.
- Incubate for 1 hour at 37°C on shaker.
- Spread 100-300 µL onto a plate made with appropriate antibiotic.
- Grow overnight at 37°C.
