Team:Groningen/protocols/Digestion
From 2013.igem.org
(Difference between revisions)
Line 64: | Line 64: | ||
<td>DNA</td> | <td>DNA</td> | ||
<td>4µl</td> | <td>4µl</td> | ||
- | <td>up to 1µ | + | <td>up to 1µg</td> |
</tr> | </tr> | ||
Line 73: | Line 73: | ||
<br> | <br> | ||
<br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf | <br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf | ||
+ | |||
+ | |||
+ | <h2>Normal digest</h2> | ||
+ | |||
+ | <h3>Materials:</h3> | ||
+ | <ul type="square"> | ||
+ | <li>MQ water</li> | ||
+ | <li>1.5ml tubes</li> | ||
+ | <li>Enzyme digestion buffer</li> | ||
+ | <li>Restriction enzymes</li> | ||
+ | <li>DNA</li> | ||
+ | </ul> | ||
+ | |||
+ | <h3>Reaction:</h3> | ||
+ | <table border="1"> | ||
+ | <tr> | ||
+ | <th>Compound</th> | ||
+ | <th>20µl</th> | ||
+ | <th>Final concentration</th> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>MQ water</td> | ||
+ | <td>up to 20µl</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>BSA</td> | ||
+ | <td>0.5µl</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>10x restriction digestion buffer</td> | ||
+ | <td>2.5µl</td> | ||
+ | <td>1x</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>restriction enzyme(s)</td> | ||
+ | <td>0.5µl</td> | ||
+ | <td>1U</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>DNA</td> | ||
+ | <td>2µl</td> | ||
+ | <td>up to 200ng</td> | ||
+ | </tr> | ||
+ | |||
+ | </table> | ||
</div> | </div> |
Revision as of 14:34, 27 July 2013
Digestion
Fast digest
Materials:
- MQ water
- 1.5ml tubes
- Fast digestion buffer
- Restriction enzymes
- DNA
Reaction:
Compound | 20µl | Final concentration |
---|---|---|
MQ water | up to 20µl | |
10x fast digestion buffer | 2µl | 1x |
restriction enzyme(s) | 1µl | 1U |
DNA | 4µl | up to 1µg |
When the reaction mix is ready, the content is mixed an incubated at 37°C for 1h.
Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf
Normal digest
Materials:
- MQ water
- 1.5ml tubes
- Enzyme digestion buffer
- Restriction enzymes
- DNA
Reaction:
Compound | 20µl | Final concentration |
---|---|---|
MQ water | up to 20µl | |
BSA | 0.5µl | |
10x restriction digestion buffer | 2.5µl | 1x |
restriction enzyme(s) | 0.5µl | 1U |
DNA | 2µl | up to 200ng |