IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/08/18

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Raphaël and Aleksandra
Test population counter (PsS/attC + PBad/Int 4))
2 conditions :

  • 1/2 dilution from the overnight culture -> 10h induction, 0,2% arabinose
  • 1/50 dilution from the overnight culture -> 10h induction, 0,2% arabinose


-> After 10h, plating with 1/100 and 1/1000 dilutions (there was just few red colonies on the plates, it's impossible to calculate the recombination rate because the constructs are in a high copy plasmid)

Léa
Minipreps

  • 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)
  • wRBS/LacZα/LuxI, 4 colonies
  • sRBS/LacZα/LuxI, 4 colonies
  • LuxI
  • pLux/LuxI, 3 colonies
  • PmW/attC, 1 colony


-> We couldn't use the nanodrop because the computer was unavailable...


Glycerols

  • 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, 2 colonies
  • 11)Ps/mRBS/LuxR + sRBS/TetA(C)/LuxI, 4 colonies (2 didn't grow)


Restriction digest with SpeI and PstI : 5µl miniprep + 15µL water + 2x1µl enzyme + 2.5µl B2 + 0.25µl BSA

  • PmW/attC


Restriction digest with XbaI and PstI : 5µl miniprep + 15µL water + 2x1µl enzyme + 2.5µl B3 + 0.25µl BSA

  • 3x Ps/wRBS/LuxR + wRBS/TetA(C)/LuxI
  • 4x Ps/wRBS/LuxR + mRBS/TetA(C)/LuxI
  • 1x Ps/sRBS/LuxR + mRBS/TetA(C)/LuxI
  • 1x Pm/sRBS/LuxR + sRBS/TetA(C)/LuxI
  • 2x Ps/wRBS/LuxR + sRBS/TetA(C)/LuxI
  • 2x Ps/mRBS/LuxR + sRBS/TetA(C)/LuxI
  • 4x wRBS/LacZα/LuxI
  • 4x sRBS/LacZα/LuxI
  • LuxI
  • 3x pLux/LuxI


Gel electrophoresis

  • Ps/wRBS/LuxR + wRBS/TetA(C)/LuxI, colony II
  • Ps/wRBS/LuxR + wRBS/TetA(C)/LuxI, colony III
  • Ps/wRBS/LuxR + wRBS/TetA(C)/LuxI, colony IV
  • Ps/wRBS/LuxR + mRBS/TetA(C)/LuxI, colony I
  • Ps/wRBS/LuxR + mRBS/TetA(C)/LuxI, colony II
  • Ps/wRBS/LuxR + mRBS/TetA(C)/LuxI, colony III
  • Ps/wRBS/LuxR + mRBS/TetA(C)/LuxI, colony IV
  • Ps/sRBS/LuxR + mRBS/TetA(C)/LuxI, colony I
  • Pm/sRBS/LuxR + sRBS/TetA(C)/LuxI, colony I
  • Ps/wRBS/LuxR + sRBS/TetA(C)/LuxI, colony I
  • Ps/wRBS/LuxR + sRBS/TetA(C)/LuxI, colony II
  • Ps/mRBS/LuxR + sRBS/TetA(C)/LuxI, colony I
  • Ps/mRBS/LuxR + sRBS/TetA(C)/LuxI, colony IV
  • pLux/LuxI, colony I
  • pLux/LuxI, colony II
  • pLux/LuxI, colony III
  • Ladder 1kb

File:Gel1808 1.jpg



Gel electrophoresis

  • PmW/attC
  • sRBS/LacZα/LuxI, colony I
  • sRBS/LacZα/LuxI, colony II
  • sRBS/LacZα/LuxI, colony III
  • sRBS/LacZα/LuxI, colony IV
  • wRBS/LacZα/LuxI, colony I
  • wRBS/LacZα/LuxI, colony II
  • wRBS/LacZα/LuxI, colony III
  • wRBS/LacZα/LuxI, colony IV
  • -
  • Ladder 1kb
  • LuxI

File:Gel1808 2.jpg