Team:Bordeaux/NotebookAugust25

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Latest revision as of 02:25, 18 October 2014

25th August

Molecular biology

  • Chewback : miniprep

  • Assembly : ELP-ELP, ELP-RLP, ELP-SLP, SLP-RLP, SLP-SLP and RLP-RLP
    • Digestion
      All OK but try again the plasmid pSB1C3 digestion

      incubation 1h30 at 37°C :
      Enzyme 1 (Nhe1 or X) 0,5μl
      Enzyme 2 (S) 0,5lμl
      10X Buffer 4 5μl
      DNA 10μl
      H2O qsp 50μl
      - Enzyme inactivated 20min at 80°C
      - Gel 1% (30min, 100V)

    • Ligation
      Plasmid 4μl
      Insert 4μl
      Buffer 10X 1μl
      Taq ligase 0,5μl
      H2O qsp 10μl
      30min at 45°C°C

    • Transformation



    Purification

    • Dialysis at 11h
    • Lyophilisation at 12h10

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