IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/08/26

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Aleksandra and Raphaël


Transformations of Turbocells

  • PmS-WTet
  • PmS-MTet
  • PsS-MTet
  • WLac-LuxI
  • SLac-LuxI
  • pLux-LuxI


Controls :

  • pLux
  • SLac
  • WLac
  • PmS
  • PsS
  • pUC19 (positive control)


-> almost no colonies on the plates (problem with Turbocells)


Colony PCR
Touchdown PCR
Primers : VF2 + VR

  • WLac-LuxI
  • PsM-MTet I
  • PsM-STet IV
  • PsS-MTet I
  • PsM-WTet I
  • PsM-MTet III
  • PsS-STet IV
  • PsS-WTet III
  • PsM-WTet II
  • PsM-STet II
  • PsS-MTet IV
  • PsS-WTet IV
  • PmS-STet I
  • PmS-WTet I
  • PmS-STet III
  • pLux-GFP (positive control)


Minipreps

  • WLac-LuxI
  • PsM-MTet I
  • PsM-STet IV
  • PsS-MTet I
  • PsM-WTet I
  • PsM-MTet III
  • PsS-STet IV
  • PsS-WTet III
  • PsM-WTet II
  • PsM-STet II
  • PsS-MTet IV
  • PsS-WTet IV
  • PmS-STet I
  • PmS-WTet I
  • PmS-STet III


Restriction digest with SpeI and EcoRI-HF : 20µl miniprep + 2.5µl B4 + 2x1µl enzymes + 0.25µl BSA

  • WLac-LuxI
  • PsM-MTet I
  • PsM-STet IV
  • PsS-MTet I
  • PsM-WTet I
  • PsM-MTet III
  • PsS-STet IV
  • PsS-WTet III
  • PsM-WTet II
  • PsM-STet II
  • PsS-MTet IV
  • PsS-WTet IV
  • PmS-STet I
  • PmS-WTet I
  • PmS-STet III


Restriction digest with XbaI and EcoRI-HF : 20µl miniprep + 2.5µl B4 + 2x1µl enzymes + 0.25µl BSA

  • I13507-pLux-GFP


Gel electrophoresis (Colony PCR)

  • WLac-LuxI
  • PsM-MTet I
  • PsM-STet IV
  • PsS-MTet I
  • PsM-WTet I
  • PsM-MTet III
  • PsS-STet IV
  • PsS-WTet III
  • PsM-WTet II
  • PsM-STet II
  • PsS-MTet IV
  • PsS-WTet IV
  • PmS-STet I
  • PmS-WTet I
  • PmS-STet III
  • pLux-GFP
  • Ladder 1Kb


Gel electrophoresis (Restriction digest)

  • PmS-STet III
  • Ladder 1Kb
  • PsM-MTet II
  • Ladder 1Kb
  • PsM-STet II
  • Ladder 1Kb
  • WLac-LuxI
  • -
  • PmS-STet I
  • -
  • PsM-MTet I
  • -


Gel electrophoresis (Restriction digest)

  • PsS-WTet III
  • Ladder 1Kb
  • PsS-MTet I
  • -
  • PsS-MTet IV
  • -
  • PsS-WTet IV
  • -
  • PsM-WTet II
  • -


Gel electrophoresis (Restriction digest)

  • PmS-WTet I
  • Ladder 1Kb
  • PsM-STet IV
  • -
  • PsM-WTet I
  • -
  • I13507-pLux-GFP


-> Electrophoresis boxes were not functioning normally and migration could not be completed


Transformations of DH5α (because Turbocells didn't work)

  • PmS-WTet
  • PmS-MTet
  • PsS-MTet
  • WLac-LuxI
  • SLac-LuxI
  • pLux-LuxI
  • PsM-MTet
  • PsM-WTet
  • PsW-MTet
  • PsW-WTet
  • PsS-WTet


Controls :

  • pLux
  • SLac
  • WLac
  • PmS
  • PsS
  • PsM
  • PsW
  • pUC19 (positive control)