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)