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