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Revision as of 08:49, 10 September 2015

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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/µlingedient
1ligation fragment (10 ng/µl)
0,5pJet
1T4 ligase
52xbuffer
2,5H2O
  • 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
,