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Team:Grenoble/Biology/Protocols/Transformation 1
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<h1>Transformation of <i>E. coli</i> competent cells (TSS method)</h1> | <h1>Transformation of <i>E. coli</i> competent cells (TSS method)</h1> | ||
<h2>Goal</h2> | <h2>Goal</h2> | ||
| - | Transform <i>E. coli</i> | + | Transform <i>E. coli</i> cells whith foreign DNA. |
</section> | </section> | ||
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Latest revision as of 19:08, 27 August 2012
Transformation of E. coli competent cells (TSS method)
Goal
Transform E. coli cells whith foreign DNA.Protocol
You need to:- Grow a liquid culture overnight.
- Dilute this culture at 1/100, in fresh LB medium.
- Let the culture grow to 0.3 OD600.
- Put 50 µL of the culture in 50 µL of cold TSS 2X.
- Incubate cells for 45 min in glaced water.
- Add 3 µL of DNA.
- Incubate cells for 10 min in glaced water.
- Incubate cells for 1.5 min at 42°C.
- Incubate cells for 10 min in glaced water.
- Add 900 µL of LB medium.
- Incubate for 1 hour at 37°C on shaker.
