Difference between revisions of "Team:Freiburg/Protocols/Blunt End Ligation"
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Latest revision as of 18:43, 16 September 2015
blunt end ligation
material: blunt end vector pjet, 2x ligation buffer, T4 Ligase, autoclaved H2O, DNA fragment of interests
duration: 7 min
- pipet the following:
| quantity/µl | ingedient |
| 1 | ligation fragment (10 ng/µl) |
| 0.5 | pJet |
| 1 | T4 ligase |
| 5 | 2xbuffer |
| 2.5 | H2O |
- incubate for 5 Min @ RT
- do a Transformation with 5 µl in E. coli TOP10, plate 100 µl
- keep the rest at 4°C as backup until transformation worked
