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Team:Alberta/Notebook/protocols/ligation
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Revision as of 00:44, 27 October 2010
Ligation
Reagents:
- Vector
- Insert
- MilliQ buffer
- 5X ligase buffer
- T4 DNA ligase
- [http://tools.invitrogen.com/content/sfs/manuals/t4dnaligase_5U_man.pdf]
Procedure:
- Volumes of vector and insert may vary, but a typical 20 ul reaction might be:
| MilliQ water | 11 ul |
| 5x ligase buffer | 4 ul |
| Vector (200 ng/ul) | 1 ul |
| Insert (200 ng/ul) | 3 ul |
| Ligase | 1 ul |
- Incubate at R/T for 1 hour or overnight at 12-16oC.
