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Team:Tuebingen/Notebook/Protocols/ligation
From 2013.igem.org
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| + | <h3>Reagents</h3> | ||
| + | <table border="0"> | ||
| + | <colgroup> | ||
| + | <col width="300"> | ||
| + | <col width="300"> | ||
| + | </colgroup> | ||
| + | <tr> | ||
| + | <td style="text-align: center">20 - 50 ng</td> | ||
| + | <td>Linearized vector</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">1 µL</td> | ||
| + | <td>10x T4 DNA Ligase Buffer</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">1 µL</td> | ||
| + | <td>T4 DNA Ligase</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">5 µl <br>(up to 5:1 molar ratio insert to vector) </td> | ||
| + | <td>Insert DNA</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">fill up to 10 µL</td> | ||
| + | <td>Aqua dest.</td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p> </p> | ||
| + | |||
| + | <h3>Procedure</h3> | ||
| + | <p>Mix all reagents and fill up with Aqua dest. to 10 µL. Vortex and incubate at 4 °C over night. On the next day, inactivate enzymes at 70 °C for 5 min. Store at -20 °C.</p> | ||
| + | |||
| + | |||
| + | <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a> | ||
</div> | </div> | ||
Latest revision as of 12:09, 4 October 2013
Ligation
Reagents
| 20 - 50 ng | Linearized vector |
| 1 µL | 10x T4 DNA Ligase Buffer |
| 1 µL | T4 DNA 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. Vortex and incubate at 4 °C over night. On the next day, inactivate enzymes at 70 °C for 5 min. Store at -20 °C.
Back to Protocols
