IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/08/17

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Aleksandra
Transformations of TURBO cells with 5µl ligation product, plating with 150µl

  • 1)Pm/sRBS/LuxR + wRBS/TetA(C)/LuxI
  • 2)Ps/wRBS/LuxR + wRBS/TetA(C)/LuxI
  • 3)Ps/mRBS/LuxR + wRBS/TetA(C)/LuxI
  • 4)Ps/sRBS/LuxR + wRBS/TetA(C)/LuxI
  • 5)Pm/sRBS/LuxR + mRBS/TetA(C)/LuxI
  • 6)Ps/wRBS/LuxR + mRBS/TetA(C)/LuxI
  • 7)Ps/mRBS/LuxR + mRBS/TetA(C)/LuxI
  • 8)Ps/sRBS/LuxR + mRBS/TetA(C)/LuxI
  • 9)Pm/sRBS/LuxR + sRBS/TetA(C)/LuxI
  • 10)Ps/wRBS/LuxR + sRBS/TetA(C)/LuxI
  • 11)Ps/mRBS/LuxR + sRBS/TetA(C)/LuxI
  • 12)Ps/sRBS/LuxR + sRBS/TetA(C)/LuxI

Controls:

  • 13)Pm/sRBS/LuxR
  • 14)Ps/wRBS/LuxR
  • 15)Ps/mRBS/LuxR
  • 16)Ps/sRBS/LuxR
  • 17)pUC19 (positive control)


Léa
Glycerols and minipreps

  • wRBS/LacZα/LuxI, 2 cultures
  • sRBS/LacZα/LuxI, 2 cultures
  • LuxI


Nanodrop of the minipreps

Sample name 260/280 260/230 ng/µl
wRBS/LacZα/LuxI cult1 1.90 2.40 422.1
wRBS/LacZα/LuxI cult2 1.95 2.34 362.3
sRBS/LacZα/LuxI cult1 1.92 2.37 387.4
sRBS/LacZα/LuxI cult2 1.94 2.35 415.2
LuxI 1.94 2.37 207.0


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

  • wRBS/LacZα/LuxI, 2 cultures
  • sRBS/LacZα/LuxI, 2 cultures
  • LuxI


Gel electrophoresis

  • Ladder 100bp
  • -
  • wRBS/LacZα/LuxI, cult1
  • -
  • wRBS/LacZα/LuxI, cult2
  • -
  • sRBS/LacZα/LuxI, cult1
  • -
  • sRBS/LacZα/LuxI, cult2
  • -
  • LuxI
  • Ladder 1kb


The restriction didn't work File:Gel1708 1.jpg


Overnight cultures

  • wRBS/LacZα/LuxI, 4 colonies
  • sRBS/LacZα/LuxI, 4 colonies
  • LuxI, 1 culture from glycerol


Raphaël
Overnight cultures

  • pLux/LuxI
  • Pm/wRBS/LuxR/attC/t/attC/mRFP
  • Ps/sRBS/LuxR/attC/t/attC/mRFP, 4 colonies


Aleksandra
Overnight cultures

  • 2)Ps/wRBS/LuxR + wRBS/TetA(C)/LuxI, 4 colonies (1 didn't grow)
  • 6)Ps/wRBS/LuxR + mRBS/TetA(C)/LuxI, 4 colonies
  • 8)Ps/sRBS/LuxR + mRBS/TetA(C)/LuxI, 1 colony
  • 9)Pm/sRBS/LuxR + sRBS/TetA(C)/LuxI, 1 colony
  • 10)Ps/wRBS/LuxR + sRBS/TetA(C)/LuxI, 4 colonies
  • 11)Ps/mRBS/LuxR + sRBS/TetA(C)/LuxI, 4 colonies (2 didn't grow)