Team:EPF Lausanne/SecondWeek

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Monday 19.07.2010

  • Results of the transformation checked: successful!!
  • Amplification of colonies (goal to do a miniprep of the plasmids --ori-- and --ori-resistance--)
  • Results of plating of asaia culture (where we hope to recover the WT) checked: Asaia cultures grew on Kan plates also! Checked colonies for fluorescance under the microscope: no fluorescance --> Very unexpected! Don't know reason yet.
  • Re-grow the colonies. Multiple cycles: hope that we get WT.
  • Preparation of Asaia cultures for Lemaitre experiments/WT/Competence...
  • Preparation of SOC medium and GLY medium
  • Autoclaving of media and flasks

Tuesday 20.07.2010

  • Plating Asaia to get WT (once again)
  • Making HEPES Buffer for the Competence of Asaia
  • Diluted the culture to get a 25ml culture(1:20) according to Asaia competence protocol
  • OD curve plotted, Doubling time calculated
  • No single colonies from the re-growing trial --> replating properly on separate plates
  • Preparation of LB medium mixed with Kan/Amp/Cm and GLY medium mixed with <kan
  • Miniprep of Origin + Resistance
  • Miniprep of Origin alone.
  • Miniprep of base vector BBa_151020
  • Concentration measurments of miniprep products
  • Glycerol stock of BB plasmid and Base Vector made

Wednesday 21.07.2010

  • Measure OD of overnight culture of Asaia for competence --> value was too high
  • Diluted culture to 0.3 OD, left in 30 degree incubator for 2 hours, OD of 0.6 measured --> start with competence
  • Asaia competence
  • Digestion of BB's produced and of the Base vector
  • Agar plates with Kan/Tet/Cm/Amp made
  • Run Gel with the products of the digestion to check if the fragments are as expected
  • Prepare Tet stock (from powder)

Thursday 22.07.2010

  • Asaia preculture for OD growth(pH) measurements
  • Gly medium with pH 2,3,4,5,6,7.

Friday 23.07.2010