IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/08/11

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Aleksandra
Transformation of TURBO cells with yesterday's ligations using 5µl ligation product to transform 50µl cells, then plating on Amp+LB with 150µl LB+transformation.

  • mRBS/TetA(C)+RBS/LuxI
  • wRBS/TetA(C)+RBS/LuxI
  • stRBS/TetA(C)+RBS/LuxI
  • stRBS/LacZα+RBS/LuxI
  • RBS/mRFP/t(K081014)+pLux/GFP
  • RBS/mRFP/tt(I13507)+pLux/GFP
  • Pm/wRBS/LuxR +attC/term/attC/mRFP
  • Ps/wRBS/LuxR +attC/term/attC/mRFP
  • Pm/mRBS/luxR +attC/term/attC/mRFP
  • Ps/mRBS/LuxR +attC/term/attC/mRFP
  • Pm/sRBS/LuxR +attC/term/attC/mRFP
  • Ps/sRBS/LuxR +attC/term/attC/mRFP


Controls:

  • RBS/LuxI
  • pLux/GFP
  • Pm/wRBS/LuxR
  • Ps/wRBS/LuxR
  • Pm/mRBS/luxR
  • Ps/mRBS/LuxR
  • Pm/sRBS/LuxR
  • Ps/sRBS/LuxR
  • Positive control (pUC19)


All the transformation plates have colonies


verification of glycerols : plating

  • wRBS/tetA(C)
  • mRBS/TetA(C)
  • stRBS/LacZα
  • Pm
  • Ps
  • Ps/wRBS/LuxR


All plates have a lot of colonies, the glycerols are OK

Théotime
Make Plate 2x 500 mL of LBA + AMP : 2x 19 plates.


Aleksandra
Starting overnight cultures, 12ml cultures LB+Amp

  • mRBS/TetA(C)+RBS/LuxI, 4 colonies
  • wRBS/TetA(C)+RBS/LuxI, 4 colonies
  • stRBS/TetA(C)+RBS/LuxI, 4 colonies
  • RBS/mRFP/t(K081014)+pLux/GFP, 4 colonies
  • RBS/mRFP/tt(I13507)+pLux/GFP, 4 colonies
  • Pm/wRBS/LuxR +attC/term/attC/mRFP, 1 possible colony, didn't grow
  • Ps/wRBS/LuxR +attC/term/attC/mRFP, 3 colonies
  • Pm/mRBS/luxR +attC/term/attC/mRFP, 1 possible colony, didn't grow
  • Ps/mRBS/LuxR +attC/term/attC/mRFP, 4 colonies
  • Pm/sRBS/LuxR +attC/term/attC/mRFP, 1 colony
  • Ps/sRBS/LuxR +attC/term/attC/mRFP, 4 colonies



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