Team:INSA-Lyon/Protocols/Transformation
From 2010.igem.org
(Difference between revisions)
m |
|||
Line 11: | Line 11: | ||
<div id="menug"> | <div id="menug"> | ||
<ul> | <ul> | ||
+ | <li><a href="/Team:INSA-Lyon/Project/Theory" class="cteal"> > Theory</a></li> | ||
+ | <li><a href="/Team:INSA-Lyon/Project/Strategy" class="slateb"> > Strategy</a></li> | ||
+ | <li><a href="/Team:INSA-Lyon/Project/Results" class="yellow"> > Results</a></li> | ||
+ | <li><a href="/Team:INSA-Lyon/Project/Future_direction" class="coral"> > Future Direction</a></li> | ||
+ | <li><a href="/Team:INSA-Lyon/Project/Notebook" class="blue"> > Notebook</a></li> | ||
+ | |||
+ | <div id="ssmenug"> | ||
<li><a href="/Team:INSA-Lyon/Project/Notebook/Calendar" class="blue"> > Calendar</a></li> | <li><a href="/Team:INSA-Lyon/Project/Notebook/Calendar" class="blue"> > Calendar</a></li> | ||
<li><a href="/Team:INSA-Lyon/Project/Notebook/Protocols" class="brn"> > Protocols</a></li> | <li><a href="/Team:INSA-Lyon/Project/Notebook/Protocols" class="brn"> > Protocols</a></li> | ||
- | <li><a href="/Team:INSA-Lyon/Project/ | + | </div> |
+ | |||
+ | <li><a href="/Team:INSA-Lyon/Project/Modeling" class="green"> > Modeling</a></li> | ||
+ | <li><a href="/Team:INSA-Lyon/Project/References" class="orange"> > References</a></li> | ||
+ | |||
</ul> | </ul> | ||
</div> | </div> |
Revision as of 10:34, 25 August 2010
To fully enjoy our wiki, we recommend you to use Firefox as web browser.
Get it now !
Thank you Groningen for sharing your code, you've been most helpful !
Protocols
Currently under construction
- 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