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Imperial College/17 August 2008
From 2008.igem.org
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17th August 2008Wet LabB.subtilisCloningMondayPrepare plates and Media for later in the week Transform E.coli Xl1-blue with C0012 (LacI) from the registry again TuesdayCheck LacI transformants Miniprep and digest 0.1.01 (Double Terminator) and 0.1.02 (Chloraphenicol Reisitance) then ligate together to form construct 0.1.14 and transform into Xl1-Blue E.coli WednesdayCheck E.coli transformed with construct 0.1.14 for growth and by PCR (if growth was succesful) If primers have arrived, PCR clone LacI, XylR, Spectinomycin, EpsE modified integration sequnce and AmyE integration sequence into Biobricks. ThursdayIf grown, mini/midi-prep a few colonies from each plate and digest a sample with XbaI and SpeI to determin which plasmids have the insert in the correct orientation. If primers arrive today PCR cloning should be carried out. FridayCheck plates from Thursday for growth and/or PCR check the minipreps that could be digested by XbaI and SpeI. |
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