User:Tk/Notebook/MF/2008/11/05

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Vector backbone

  • gave up on biotin purification
  • PCR cleanup after PCR with plain primers (62 C anneal) 100 ul rxn
  • elute 2x with 40 ul in EB
  • digestion in 100 ul with E/P 2 hr / 20 min heat kill
  • PCR cleanup, elute 2x with 40 ul
  • yields 1AC3: 48 1AK3: 69 1AT3: 57 ng/ul


Transformation results

  • one colony on the AC plate, none on others.
  • control normal at 4e7 efficiency
  • culture the colony in Amp/Kan and Amp/Tet medium