IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/09/09

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Minipreps

  • PmS-MTet-LuxI
  • LuxI
  • attP
  • PsS-RFP
  • PsW-RFP
  • PsM-RFP
  • PmS-RFP
  • pBad/Int
  • Final PmS-MTet x5
  • LuxI/GFP x1
  • pLux/LuxI x1


Restriction digest


With XbaI and PstI

  • LuxI/GFP
  • I13507/pLux/GFP
  • RBS-GFP-tt (E0240)


With SpeI and PstI

  • PmS-MTet-LuxI
  • LuxI
  • pLux/LuxI
  • pLux


With EcoRI-HF and PstI

  • PmS-MTet-LuxI
  • PsS-RFP
  • PsW-RFP
  • PsM-RFP
  • PmS-RFP
  • attP
  • Final PmS-MTet x5


Gel electrophoresis

  • PmS-MTet-LuxI (E+P)
  • PsS-RFP
  • PsW-RFP
  • PsM-RFP
  • PmS-RFP
  • attP
  • I13507/pLux/GFP
  • Final PmS-MTet 1
  • Final PmS-MTet 2
  • Final PmS-MTet 3
  • Final PmS-MTet 4
  • Final PmS-MTet 5
  • LuxI/GFP
  • RBS-GFP-tt (E0240)


Ligations (V + I)

  • 1) Ps + WTet
  • 2) Ps + MTet
  • 3) Ps + STet
  • 4) Ctrl Ps


Make competent cells

  • Turbocells (100mL of culture)
  • PmS Chr (to transform with pBad/Int)


Transformation of 1-4 + positive control (pUC19) for new Turbocells


Overnight cultures

  • Plambda Int


From transformations of yesterday :

  • PsS-WTet-LuxI + pBad/Int x2
  • PmS-MTet-LuxI + pBad/Int x 2