IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/09/12

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Glycerols

  • Ps-MTet x2
  • Ps-STet x2
  • PsS-WTet-LuxI 1 and 2
  • 6h A+ PmS
  • 5h A+ psS
  • 6h ctrl A PmS
  • 5h ctrl A PsS
  • 5h ctrl A PmS
  • 4h ctrl A PsS


Minipreps

  • 6h A+ PmS
  • 5h A+ psS
  • 6h ctrl A PmS
  • 5h ctrl A PsS
  • 5h ctrl A PmS
  • 4h ctrl A PsS
  • PsS-WTet-LuxI 1 and 2
  • Ps-MTet 1 and 2
  • Ps-STet 1 and 2


Gel electrophoresis with PCR from 09/10
many bands on the gel but the 4Kb band is seen (PmS integrated into the chromosome ?)


PCR to verify that the terminator is out in our A+ constructs (primers : VF2 + VR)

  • 6h A+ PmS
  • 5h A+ psS
  • 6h ctrl A PmS
  • 5h ctrl A PsS
  • 5h ctrl A PmS
  • 4h ctrl A PsS


Gel electrophoresis

  • 6h A+ PmS
  • 5h A+ psS
  • 6h ctrl A PmS
  • 5h ctrl A PsS
  • 5h ctrl A PmS
  • 4h ctrl A PsS


strange results


Ligations (V + I)

  • 1) PmS-MTet-LuxI + I13507/pLux/GFP
  • 2) pLux/LuxI + RBS-GFP-tt (E0240)
  • 3) pLux + LuxI
  • 4) Ctrl PmS-MTet-LuxI
  • 5) Ctrl pLux/LuxI
  • 6) Ctrl pLux


Transformation of 1-6


Overnight cultures

  • PmS Chr + pBad/Int x2



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