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Protocol/8
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Revision as of 00:35, 26 October 2010
Protocol 8: Ligation
What you will need:
- Vector
- Insert
- Milli Q buffer
- 5X ligase buffer
- T4 DNA ligase
- [http://tools.invitrogen.com/content/sfs/manuals/t4dnaligase_5U_man.pdf]
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 hrs or overnight at 12-16 C
