IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/08/20

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

  • Left arm HK022 Mut-Mut
  • Left arm HK022 WT-Mut
  • Left arm λ Mut-Mut
  • Left arm λ WT-Mut
  • Left arm WT-WT


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

  • Kan resistance cassette


Restriction digest with XbaI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B3 + 0.25µl BSA

  • Right arm HK022 Mut-Mut
  • Right arm HK022 WT-Mut
  • Right arm λ Mut-Mut
  • Right arm λ WT-Mut


Restriction digest with SpeI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B2 + 0.25µl BSA

  • LacZα expression cassette
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Aleksandra
Minipreps

  • Ps/wRBS/LuxR + attC/term/attC/mRFP
  • Ps/mRBS/LuxR + attC/term/attC/mRFP
  • Ps/sRBS/LuxR + attC/term/attC/mRFP
  • Pm/sRBS/LuxR + attC/term/attC/mRFP
  • pLux
  • wRBS/LacZ
  • sRBS/LacZ
  • wRBS/TetA(C)
  • mRBS/TetA(C)
  • 2x RBS/LuxI


Raphaël
Restriction digest with XbaI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B2 + 0.25µl BSA

  • wRBS/TetA(C)
  • mRBS/TetA(C)
  • 2x RBS/LuxI


Restriction digest with SpeI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B2 + 0.25µl BSA

  • Ps/wRBS/LuxR + attC/term/attC/mRFP
  • Ps/mRBS/LuxR + attC/term/attC/mRFP
  • Ps/sRBS/LuxR + attC/term/attC/mRFP
  • Pm/sRBS/LuxR + attC/term/attC/mRFP
  • pLux
  • wRBS/LacZ
  • sRBS/LacZ


Aleksandra and Raphaël


Gel electrophoresis

  • Ps/wRBS/LuxR + attC/term/attC/mRFP
  • Ladder 1kb
  • Ps/mRBS/LuxR + attC/term/attC/mRFP
  • -
  • Ps/sRBS/LuxR + attC/term/attC/mRFP
  • -
  • Pm/sRBS/LuxR + attC/term/attC/mRFP
  • -
  • pLux
  • -
  • wRBS/LacZ


File:Gel2008 1.jpg


Gel electrophoresis

  • sRBS/LacZ
  • -
  • wRBS/TetA(C)
  • -
  • mRBS/TetA(C)
  • -
  • RBS/LuxI
  • RBS/LuxI
  • Ladder 1kb
  • -
  • -
  • -


-> The gel was unfortunately damaged... thus we have lost a lot of DNA and the migration was abnormal.


File:Gel2008 2.jpg

Léa
Gel extraction

  • Ps/sRBS/LuxR + attC/term/attC/mRFP
  • Pm/sRBS/LuxR + attC/term/attC/mRFP
  • pLux
  • wRBS/LacZ