Team:INSA-Lyon/Protocols/Digestio
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- | < | + | <div id="menug"> |
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+ | <li><a href="/Team:INSA-Lyon/Notebook/Calendar"> > Calendar</a></li> | ||
+ | <li><a href="/Team:INSA-Lyon/Notebook/Protocols" <!-- class="green" --> > Protocols</a></li> | ||
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+ | <p style="text-align:justify ; color:white;"><a href="http://www.mozilla.com/en-US/"><img style="width:20px; height:20px;" src="http://icones.eee-pc.fr/icones/firefox3.png" alt="firefox" /></a> To fully enjoy our wiki, we recommend you to use Firefox as web browser. | ||
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+ | |||
+ | <br> | ||
+ | <br> | ||
+ | <br><em>Thank you Groningen for sharing your code, you've been most helpful !</em><br><br></p> | ||
+ | </div> | ||
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+ | <div id="corps"> | ||
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+ | <h3> Protocols </h3> | ||
+ | <p>Currently under construction <br><br><br><br></p> | ||
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+ | <h5 style="text-indent:20px">Digestion</h5> | ||
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+ | <h5>Single digestion :</h5> | ||
+ | <br><br> | ||
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<li>Add x µL DNA (0,5 to 1 ug)</li> | <li>Add x µL DNA (0,5 to 1 ug)</li> | ||
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<li>Incubate 1h at 37°C.</li> | <li>Incubate 1h at 37°C.</li> | ||
</ol> | </ol> | ||
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- | < | + | <h5>Double digestion EcoRI/SpeI:</h5> |
+ | <br><br> | ||
<ol id="list_extra"> | <ol id="list_extra"> | ||
<li>Add x µL DNA (0,5 to 1 µg). Add 15 µL DNA extracted from the part transformation.</li> | <li>Add x µL DNA (0,5 to 1 µg). Add 15 µL DNA extracted from the part transformation.</li> | ||
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<li>Incubate 1h at 37°C</li> | <li>Incubate 1h at 37°C</li> | ||
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- | + | <h5>Double digestion EcoRI/XbaI :</h5> | |
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<li>Add x µL DNA. Add 15 µL DNA extracted from the part transformation.</li> | <li>Add x µL DNA. Add 15 µL DNA extracted from the part transformation.</li> | ||
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<li>Incubate 1h at 37°C</li> | <li>Incubate 1h at 37°C</li> | ||
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- | + | <h5>Double digestion XbaI/PstI :</h5> | |
- | < | + | <br><br> |
<ol id="list_extra"> | <ol id="list_extra"> | ||
<li>Add x µL DNA. Add 15 µL DNA extracted from the part transformation.</li> | <li>Add x µL DNA. Add 15 µL DNA extracted from the part transformation.</li> | ||
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<li>Incubate 1h at 37°C</li> | <li>Incubate 1h at 37°C</li> | ||
</ol> | </ol> | ||
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- | + | ||
- | < | + | <h5>Double digestion XbaI/SpeI :</h5> |
+ | <br><br> | ||
<ol id="list_extra"> | <ol id="list_extra"> | ||
<li>Add x µL DNA. Add 15 µL DNA extracted from the part transformation.</li> | <li>Add x µL DNA. Add 15 µL DNA extracted from the part transformation.</li> | ||
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<li>Incubate 1h at 37°C </li> | <li>Incubate 1h at 37°C </li> | ||
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Revision as of 14:44, 23 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
Digestion
Single digestion :
- Add x µL DNA (0,5 to 1 ug)
- Add 15 µL DNA extracted from the part transformation.
- Add 2 µL 10X buffer
- Add the enzyme last
- Add 0,5 to 2 µL restriction enzyme (max 1/10 of final volume)
- Add sterile water qs 20 µL
- Incubate 1h at 37°C.
Double digestion EcoRI/SpeI:
- Add x µL DNA (0,5 to 1 µg). Add 15 µL DNA extracted from the part transformation.
- Add 2 µL buffer Tango 10x
- Add 1 µL SpeI
- Add sterile water qs 20 µL
- Incubate 1h at 37°C
- Add 2 µL buffer Tango 10x
- Add 1 µL EcoRI
- Incubate 1h at 37°C
Double digestion EcoRI/XbaI :
- Add x µL DNA. Add 15 µL DNA extracted from the part transformation.
- Add 4 µL buffer Tango 10x
- Add 1 µL XbaI
- Add 0,5 µL EcoRI
- Add sterile water qs 20 µL
- Incubate 1h at 37°C
Double digestion XbaI/PstI :
- Add x µL DNA. Add 15 µL DNA extracted from the part transformation.
- Add 2 µL buffer Tango 10x
- Add 1µL XbaI
- Add 1 µL PstI
- Add sterile water qs 20 µL
- Incubate 1h at 37°C
Double digestion XbaI/SpeI :
- Add x µL DNA. Add 15 µL DNA extracted from the part transformation.
- Add 2 µL buffer Tango 10x
- Add 1 µL XbaI
- Add 1 µL SpeI
- Add sterile water qs 20 µL
- Incubate 1h at 37°C