IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/06/30

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Results of Competent Cells Testing

  • The DH5α A and B stocks and DB3.1 cells all had red colonies that were too numerous to count
  • All cells are competent, the few cell count on previous testing was likely due to the pUC19 plasmid
  • Glycerol stocks of A and B stocks DH5α cells were made by:
    • Adding 0.618mL 100% glycerol to 3.8mL competent cells to make a 14% glycerol solution
    • Store in -80°C freezer

Transformation of pBADwt+RFP and pBADweak+RFP into BW27783

  • As pBADwt+RFP and pBADweak+RFP are currently just in DH5α glycerol stocks, need to transfer them to BW27783 cells which will have homogenous induction in the population upon stimulation with arabinose
  • The plasmids were transformed using the iGEM:British Columbia Transfromation protocol with 3μL of miniprepped pBADwt+RFP and pBADweak+RFP
  • A control transformation performed under the same conditions except for 1μL of 5ng/μL J23100 miniprepped plasmid was used
  • J23100 cells were plated on Amp
  • pBADwt+RFP were plated on Chlor
  • pBADweak+RFP was plated on Amp-Tet