IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/09/08

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Ligations (V + I)

  • 1) pLux + LuxI
  • 2) RBS/GFP/tt (E0240) + LuxI
  • 3) Pms/mTet/LuxI + I13507/pLux/GFP
  • 4) Ctrl pLux
  • 5) Ctrl RBS/GFP/tt (E0240)
  • 6) Ctrl Pms/mTet/LuxI


Transformation of 1-6 + positive control


Minipreps

  • PmS-MTet-LuxI
  • WTet
  • MTet
  • STet
  • Ps
  • pSB1C3 PmS-MTet-LuxI I
  • pSB1C3 PsS-WTet-LuxI1 I
  • PsS-WTet-LuxI2
  • PsS-WTet-LuxI1
  • pSB1C3 PsS-WTet-LuxI2 I
  • pSB1C3 PmS-MTet-LuxI II


Restriction digest


With XbaI and PstI

  • WTet
  • MTet
  • STet


With SpeI and PstI

  • Ps


With EcoRI-HF and SpeI

  • pSB1C3 PmS-MTet-LuxI I
  • pSB1C3 PsS-WTet-LuxI1 I
  • pSB1C3 PsS-WTet-LuxI2 I
  • pSB1C3 PmS-MTet-LuxI II
  • PmS-MTet-LuxI


Gel electrophoresis

  • WTet
  • MTet
  • STet
  • pSB1C3 PmS-MTet-LuxI I
  • pSB1C3 PsS-WTet-LuxI1 I
  • pSB1C3 PsS-WTet-LuxI2 I
  • pSB1C3 PmS-MTet-LuxI II
  • PmS-MTet-LuxI


Gel extraction

  • WTet
  • MTet
  • STet


PCR purification

  • Ps


Transformation

  • PsS-WTet + pBad/Int
  • PmS-MTet + pBad/Int
  • positive control


Overnight cultures

  • PmS-MTet-LuxI
  • PsS-WTet-LuxI
  • PmS Chr
  • LuxI
  • attP
  • PsS-RFP
  • PsW-RFP
  • PsM-RFP
  • PmS-RFP
  • pBad/Int


From transformations of this morning :

  • Final PmS-MTet x5
  • LuxI/GFP x1
  • pLux/LuxI x2 -> one colony didnt' grow