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Team:Tuebingen/Notebook/Protocols/ligation
From 2013.igem.org
(Difference between revisions)
| Line 48: | Line 48: | ||
<tr> | <tr> | ||
| - | <td style="text-align: center">to 10 µL</td> | + | <td style="text-align: center">fill up to 10 µL</td> |
<td>Aqua dest.</td> | <td>Aqua dest.</td> | ||
</tr> | </tr> | ||
| Line 56: | Line 56: | ||
<h3>Procedure</h3> | <h3>Procedure</h3> | ||
| - | <p> | + | <p>Mix all reagents and fill up with Aqua dest. to 10 µL. Incubate at 4 °C over night.</p> |
| + | |||
| + | <p>On the next day, inactivate enzymes at 70 °C for 5 min.</p> | ||
</div> | </div> | ||
Revision as of 00:23, 4 October 2013
Ligation
Back to Protocols
Reagents
| 20 - 50 ng | Linearized vector |
| 1 µL | 10x Ligase buffer |
| 1 µL | T4 Ligase |
| 5 µl (up to 5:1 molar ratio insert to vector) |
Insert DNA |
| fill up to 10 µL | Aqua dest. |
Procedure
Mix all reagents and fill up with Aqua dest. to 10 µL. Incubate at 4 °C over night.
On the next day, inactivate enzymes at 70 °C for 5 min.
