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Team:Groningen/protocols/Digestion
From 2013.igem.org
(Difference between revisions)
| 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 39: | Line 39: | ||
<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 ALIGN=CENTER>up to 20µl</td> | + | <td ALIGN=CENTER>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 ALIGN=CENTER>2µl</td> | + | <td ALIGN=CENTER>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 ALIGN=CENTER>1µl</td> | + | <td ALIGN=CENTER>1 µl</td> |
<td>1U</td> | <td>1U</td> | ||
</tr> | </tr> | ||
| Line 63: | Line 63: | ||
<tr> | <tr> | ||
<td>DNA</td> | <td>DNA</td> | ||
| - | <td ALIGN=CENTER>4µl</td> | + | <td ALIGN=CENTER>4 µl</td> |
| - | <td>up to 1µg</td> | + | <td>up to 1 µg</td> |
</tr> | </tr> | ||
| Line 81: | Line 81: | ||
<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 91: | Line 91: | ||
<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 97: | ||
<tr> | <tr> | ||
<td>MQ water</td> | <td>MQ water</td> | ||
| - | <td ALIGN=CENTER>up to 20µl</td> | + | <td ALIGN=CENTER>up to 20 µl</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
| Line 103: | Line 103: | ||
<tr> | <tr> | ||
<td>10x restriction digestion buffer</td> | <td>10x restriction digestion buffer</td> | ||
| - | <td ALIGN=CENTER>2µl</td> | + | <td ALIGN=CENTER>2 µl</td> |
<td>1x</td> | <td>1x</td> | ||
</tr> | </tr> | ||
| Line 109: | Line 109: | ||
<tr> | <tr> | ||
<td>restriction enzyme(s)</td> | <td>restriction enzyme(s)</td> | ||
| - | <td ALIGN=CENTER>0.5µl</td> | + | <td ALIGN=CENTER>0.5 µl</td> |
<td>1U</td> | <td>1U</td> | ||
</tr> | </tr> | ||
| Line 115: | Line 115: | ||
<tr> | <tr> | ||
<td>DNA</td> | <td>DNA</td> | ||
| - | <td ALIGN=CENTER>2µl</td> | + | <td ALIGN=CENTER>2 µl</td> |
| - | <td>0.2-1.5µg</td> | + | <td>0.2-1.5 µg</td> |
</tr> | </tr> | ||
Revision as of 15:03, 27 September 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.
Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf
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.
iGEM 2013 Groningen
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