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Team:Groningen/protocols/Digestion
From 2013.igem.org
(Difference between revisions)
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<div class="mainContent"> | <div class="mainContent"> | ||
| - | < | + | <h2>Restriction digestion</h2> |
<h3>Fast digest</h3> | <h3>Fast digest</h3> | ||
| Line 29: | Line 29: | ||
<ul type="square"> | <ul type="square"> | ||
<li>MQ water</li> | <li>MQ water</li> | ||
| - | <li>1. | + | <li>1.5 ml tubes</li> |
<li>Fast digestion buffer</li> | <li>Fast digestion buffer</li> | ||
<li>Restriction enzymes</li> | <li>Restriction enzymes</li> | ||
| Line 36: | Line 36: | ||
<h5>Reaction:</h5> | <h5>Reaction:</h5> | ||
| - | <table | + | <table id="normal" width="60%"> |
<tr> | <tr> | ||
<th>Compound</th> | <th>Compound</th> | ||
| - | <th>20µl</th> | + | <th>20 µl</th> |
<th>Final concentration</th> | <th>Final concentration</th> | ||
</tr> | </tr> | ||
| Line 45: | Line 45: | ||
<tr> | <tr> | ||
<td>MQ water</td> | <td>MQ water</td> | ||
| - | <td | + | <td>up to 20 µl</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
| Line 51: | Line 51: | ||
<tr> | <tr> | ||
<td>10x fast digestion buffer</td> | <td>10x fast digestion buffer</td> | ||
| - | <td | + | <td>2 µl</td> |
<td>1x</td> | <td>1x</td> | ||
</tr> | </tr> | ||
| Line 57: | Line 57: | ||
<tr> | <tr> | ||
<td>restriction enzyme(s)</td> | <td>restriction enzyme(s)</td> | ||
| - | <td | + | <td>1 µl</td> |
<td>1U</td> | <td>1U</td> | ||
</tr> | </tr> | ||
| Line 63: | Line 63: | ||
<tr> | <tr> | ||
<td>DNA</td> | <td>DNA</td> | ||
| - | <td | + | <td>4 µl</td> |
| - | <td>up to 1µg</td> | + | <td>up to 1 µg</td> |
</tr> | </tr> | ||
| Line 70: | Line 70: | ||
| - | <br>When the reaction mix is ready, the content is mixed and incubated at 37°C for 1h. | + | <br>When the reaction mix is ready, the content is mixed and incubated at 37°C for 1h [1]. |
| - | + | ||
| - | + | ||
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<ul type="square"> | <ul type="square"> | ||
<li>MQ water</li> | <li>MQ water</li> | ||
| - | <li>1. | + | <li>1.5 ml tubes</li> |
<li>Enzyme digestion buffer</li> | <li>Enzyme digestion buffer</li> | ||
<li>Restriction enzymes</li> | <li>Restriction enzymes</li> | ||
| Line 88: | Line 86: | ||
<h5>Reaction:</h5> | <h5>Reaction:</h5> | ||
| - | <table | + | <table id="normal" width="60%"> |
<tr> | <tr> | ||
<th>Compound</th> | <th>Compound</th> | ||
| - | <th>20µl</th> | + | <th>20 µl</th> |
<th>Final concentration</th> | <th>Final concentration</th> | ||
</tr> | </tr> | ||
| Line 97: | Line 95: | ||
<tr> | <tr> | ||
<td>MQ water</td> | <td>MQ water</td> | ||
| - | <td | + | <td>up to 20 µl</td> |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
<td></td> | <td></td> | ||
</tr> | </tr> | ||
| Line 109: | Line 101: | ||
<tr> | <tr> | ||
<td>10x restriction digestion buffer</td> | <td>10x restriction digestion buffer</td> | ||
| - | <td | + | <td>2 µl</td> |
<td>1x</td> | <td>1x</td> | ||
</tr> | </tr> | ||
| Line 115: | Line 107: | ||
<tr> | <tr> | ||
<td>restriction enzyme(s)</td> | <td>restriction enzyme(s)</td> | ||
| - | <td | + | <td>0.5 µl</td> |
<td>1U</td> | <td>1U</td> | ||
</tr> | </tr> | ||
| Line 121: | Line 113: | ||
<tr> | <tr> | ||
<td>DNA</td> | <td>DNA</td> | ||
| - | <td | + | <td>2 µl</td> |
| - | <td>0.2-1.5µg</td> | + | <td>0.2-1.5 µg</td> |
</tr> | </tr> | ||
| Line 129: | Line 121: | ||
<br>When the reaction mix is ready, the content is mixed and incubated at 37°C for 2h. | <br>When the reaction mix is ready, the content is mixed and incubated at 37°C for 2h. | ||
<br> | <br> | ||
| - | + | <br> [1] Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf | |
</div> | </div> | ||
Latest revision as of 23:32, 4 October 2013
Restriction digestion
Fast digest
Materials:
- MQ water
- 1.5 ml tubes
- Fast digestion buffer
- Restriction enzymes
- DNA
Reaction:
| Compound | 20 µl | Final concentration |
|---|---|---|
| MQ water | up to 20 µl | |
| 10x fast digestion buffer | 2 µl | 1x |
| restriction enzyme(s) | 1 µl | 1U |
| DNA | 4 µl | up to 1 µg |
When the reaction mix is ready, the content is mixed and incubated at 37°C for 1h [1].
Normal digest
Materials:
- MQ water
- 1.5 ml tubes
- Enzyme digestion buffer
- Restriction enzymes
- DNA
Reaction:
| Compound | 20 µl | Final concentration |
|---|---|---|
| MQ water | up to 20 µl | |
| 10x restriction digestion buffer | 2 µl | 1x |
| restriction enzyme(s) | 0.5 µl | 1U |
| DNA | 2 µl | 0.2-1.5 µg |
When the reaction mix is ready, the content is mixed and incubated at 37°C for 2h.
[1] Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf
iGEM 2013 Groningen
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