Team:INSA-Lyon/Protocols/Digestio
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+ | <div id="corps2"> | ||
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+ | <h3> Protocols </h3> | ||
+ | <br><p>Choose a protocol to read its description : | ||
+ | <br> | ||
+ | <br></p> | ||
+ | </html> | ||
{{Template:INSALyon2010_Protocole}} | {{Template:INSALyon2010_Protocole}} | ||
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- | <html | + | <br><br><br><br><br> |
+ | <h5 style="text-indent:20px">Digestion</h5> | ||
+ | <br><br> | ||
+ | |||
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- | < | + | |
+ | <br><br><br> | ||
- | |||
- | |||
+ | <h5>Single digestion :</h5> | ||
+ | <br><br> | ||
<ol id="list_extra"> | <ol id="list_extra"> | ||
<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> | ||
- | < | + | <br><br><br><br><br> |
- | + | ||
- | < | + | <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> | ||
</ol> | </ol> | ||
- | < | + | <br><br><br><br><br> |
+ | |||
- | + | <h5>Double digestion EcoRI/XbaI :</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> | ||
- | < | + | <br><br><br><br><br> |
+ | |||
- | + | <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> | ||
- | < | + | <br><br><br><br><br> |
- | + | ||
- | < | + | <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> | ||
</ol> | </ol> | ||
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+ | |||
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+ | </div> | ||
</html> | </html> |
Latest revision as of 21:09, 25 October 2010
Protocols
Choose a protocol to read its description :
- 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