IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/09/01

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Raphaël
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
  • wRBS/TetA(C)/LuxI
  • mRBS/TetA(C)/LuxI
  • sRBS/TetA(C)/LuxI
  • wRBS/LacZα
  • sRBS/LacZα
  • LuxI
  • I13507/pLux/GFP
  • pLux
  • attP (in CmR)


Restriction digest
15µl DNA + 2x1µl enzymes + 2µl buffer + 0.2µl BSA
With SpeI and PstI (B2):

  • 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
  • wRBS/LacZα
  • sRBS/LacZα
  • pLux


With XbaI ansd PstI (B3):

  • wRBS/TetA(C)/LuxI
  • mRBS/TetA(C)/LuxI
  • sRBS/TetA(C)/LuxI
  • LuxI


With EcoRI-HF and SpeI (B4):

  • I13507/pLux/GFP


Gel electrophoresis
Gel 1

  • 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
  • -
  • -
  • Ladder 1kb
  • -
  • -

File:Gel0109 1.jpg


Gel 2

  • wRBS/TetA(C)/LuxI
  • -
  • mRBS/TetA(C)/LuxI
  • -
  • sRBS/TetA(C)/LuxI
  • -
  • LuxI
  • -
  • I13507/pLux/GFP
  • Ladder 1 kb
  • -

File:Gel0109 2.jpg


Gel extraction

  • Ps/mRBS/LuxR/attC/term/attC/mRFP
  • Ps/sRBS/LuxR/attC/term/attC/mRFP
  • Pm/sRBS/LuxR/attC/term/attC/mRFP
  • LuxI


PCR purification kit

  • wRBS/LacZα
  • sRBS/LacZα
  • pLux


Restriction digest
5µl DNA + 10µl water + 2x1µl enzymes + 2µl buffer + 0.2µl BSA
With SpeI and PstI (B2):

  • 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


With EcoRI-HF and XbaI (B4):

  • I13507/pLux/GFP


With XbaI and PstI and AlWNI (B2):

  • wRBS/TetA(C)/LuxI
  • mRBS/TetA(C)/LuxI
  • sRBS/TetA(C)/LuxI


Gel electrophoresis

  • -
  • 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
  • -
  • I13507/pLux/GFP
  • -
  • wRBS/TetA(C)/LuxI
  • -
  • mRBS/TetA(C)/LuxI
  • -
  • sRBS/TetA(C)/LuxI
  • Ladder 1kb

File:Gel0109 3.jpg


Overnight cultures

  • Pm/sRBS/LuxR/attC/term/attC/mRFP
  • attP-E (pSWK)