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Team:Tuebingen/Notebook/Protocols/restriction
From 2014.igem.org
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<h3>Preparative restriction digests</h3> | <h3>Preparative restriction digests</h3> | ||
<h4>Reagents</h4> | <h4>Reagents</h4> | ||
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<td style="text-align: center">to 100 µL</td> | <td style="text-align: center">to 100 µL</td> | ||
| - | <td> | + | <td>H<sub>2</sub>O</td> |
</tr> | </tr> | ||
</table> | </table> | ||
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<h4>Procedure</h4> | <h4>Procedure</h4> | ||
Latest revision as of 15:34, 17 October 2014
Protocols
Preparative restriction digests
Reagents
| min. 2 µg | Plasmid DNA |
| 2 µL | of each restriction enzyme |
| 10 µL | 10x NE buffer 2 |
| 1 µL | 100x BSA |
| to 100 µL | H2O |
Procedure
- After mixing the reagents in a 1.5 ml Eppendorf-tube: Incubation at 37 °C overnight.
- Heat inactivate the restriction enzymes at 80 °C for 20 min.
- Treat the plasmid backbones with Antarctic Phosphatase (NEB-protocol).
- Separation via gel electrophoresis and gel extraction.
