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Team:Tuebingen/Notebook/Protocols/ligation
From 2014.igem.org
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Revision as of 23:10, 14 October 2014
DNA-Ligation
Reagents
| 20-50 ng | linearized vector |
| 1 µL | 10x ligation buffer |
| 1 µL | T4 ligase |
| 1 µL | T4 ligase |
| 1 µL | ATP |
| 100-200 ng | Insert |
| to 10 µL | H20 |
Procedure
- Mix all reagents in a PCR-tube and if necessary add to 10 μl with H20.
- Run cycler starting at 25 °C, reducing temperature hourly by 1 degree and remaining at 4 °C.
- Heat inactivate the enzymes for 5 min at 70 °C.
- Store at -20 °C.
