IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/06/29

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Testing Competent Cells

  • Competent cell stocks A and B and a DB3.1 aliquot were transformed with J23100+RBS using the iGEM:British Columbia transformation protocol to determine if the cells are competent

Testing pBAD weak+RFP and pBAD wt + RFP

  • To test if the mutation of pBAD will work, both strains are grown in three liquid cultures containing LB+antibiotic and 0.01% arabinose, 0.1% arabinose and 1% arabinose
  • After 3 hours, no red was seen in the pBAD weak+RFP strain. All three cultures of pBADwt started to express a similar level of RFP that could be seen with the human eye.
  • After 5.5hours all three cultures of pBADwt showed a stronger color of red. The 1% and 0.1% arabinose cultures of pBAD weak expressed some RFP but at lower levels then pBADwt, 0.01% arabinose culture did not show any red that could be seen
  • After 12 hours pBADwt was not significantly darker in color. pBAD weak exprssed more RFP, but was still considerably lighter then the pBAdwt strain

NOTE: pBADwt+RFP was in BW27783 cells and pBADweak+RFP was in DH5a cells, this may have contributed to the color variation.


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