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Team:TU Darmstadt/Notebook/Methods/DNA ligation
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Revision as of 16:07, 16 October 2014
DNA ligation
Short explanation:
DNA ligation is necessary to assemble digested DNA parts into a vector. The cut ends generated by restriciton enzymes are put together by DNA ligase.
Procedure: 20 µL batch
- T4 Ligase Buffer 2 µL
- T4 Ligase 1 µL
- digested vectors 25 ng (Molar ratio = Insert : backbone 3:1)
- add ddH2O to 20µL
- Incubate at 16°C overnight, alternatively incubate at room temperature for 30 min (results might be worse)
- Inactivate 20min at 80°C
