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Preparation of freezable competent cells (M15): 40 alicos of 100µL Test of the competent cells.
Begining of ‘intra-cell ROS concentration’ modelisation. Transfection of competent cells with the biobricks : - pLac - mCherry - cl Inverter - RBS (Elowitz 1999) - GFP First contact with Arduino : the electronic device that will be used as a controler, just as the Cambridge 2010 iGEM team. Culture of the BBs k174000 and K592004/5/6
Failure with the transfections. Other problem with the petri dishes : everything grew, even the WT colonies that should have died because of antibiotics. Maybe the antibiotics are too old since they were used by Grenoble iGEM Team of 2012
Preparation of freezable competent cells (WT)(40 alicos of 100µL) Same transfections than wednesday, with fresh antibiotics. PCR on pAraBad, mCherry and KillerRed, only pAraBAD worked. Team meeting TODO List : - Find a name - Begin construction of : ->pLac-RBS-YF1-fixJ ->pFixK2-RBS-GFP - glycerol-stock BBs - E-glometer, caracterisation of LED
Little Summary with Keywords
Incubated a 50mL overnight culture of E. coli M15[pRep4] cells (Qiagen, Venlo, Netherlands), containing the pQE30::αSNAP vector, in preparation for midiprep.
Midi-prepped pQE30::αSNAP. The DNA sample concentration was only of 10 ng/µL. The experiment has to be re performed.
Incubated a 10mL overnight culture of E. coli M15[pRep4-pQE30::αSNAP] cells, in preparation for miniprep.
Mini-prepped pQE30::αSNAP/pRep4 and got a 98,9 ng/µL DNA sample.
Digested pQE30::αSNAP with BamHI and KpnI restriction enzymes. Separation of the gene of non interest (αSNAP) from the pQE30 vector backbone by gel electrophoresis (1.2 % agarose, 30 min, 135V).
