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Team:Groningen/protocols/Digestion
From 2013.igem.org
(Difference between revisions)
| Line 97: | Line 97: | ||
<tr> | <tr> | ||
<td>MQ water</td> | <td>MQ water</td> | ||
| - | <td>up to 20µl</td> | + | <td ALIGN=CENTER>up to 20µl</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
| Line 103: | Line 103: | ||
<tr> | <tr> | ||
<td>BSA</td> | <td>BSA</td> | ||
| - | <td>0.5µl</td> | + | <td ALIGN=CENTER>0.5µl</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
| Line 109: | Line 109: | ||
<tr> | <tr> | ||
<td>10x restriction digestion buffer</td> | <td>10x restriction digestion buffer</td> | ||
| - | <td>2µl</td> | + | <td ALIGN=CENTER>2µl</td> |
<td>1x</td> | <td>1x</td> | ||
</tr> | </tr> | ||
| Line 115: | Line 115: | ||
<tr> | <tr> | ||
<td>restriction enzyme(s)</td> | <td>restriction enzyme(s)</td> | ||
| - | <td>0.5µl</td> | + | <td ALIGN=CENTER>0.5µl</td> |
<td>1U</td> | <td>1U</td> | ||
</tr> | </tr> | ||
| Line 121: | Line 121: | ||
<tr> | <tr> | ||
<td>DNA</td> | <td>DNA</td> | ||
| - | <td>2µl</td> | + | <td ALIGN=CENTER>2µl</td> |
<td>0.2-1.5µg</td> | <td>0.2-1.5µg</td> | ||
</tr> | </tr> | ||
Revision as of 14:48, 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 | 2µl | 1x |
| restriction enzyme(s) | 0.5µl | 1U |
| DNA | 2µl | 0.2-1.5µg |
When the reaction mix is ready, the content is mixed an incubated at 37°C for 2h.
iGEM 2013 Groningen
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