Revision as of 03:27, 27 October 2012 by Cmwinter (Talk | contribs)

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Escaper assay for the rhamnose system

The following assay is based on CFU in both Top 10 and glyA knockout of E. coli.

  • The glyA auxotroph was grown in LB media overnight at 37oC with shaking.
  • Cells were washed in M9 minimal media two times and spun in between at 4,000 RPM at 4oC for 10 min. each time.
  • Finally, washed cells were subcultured into 5 mL of M9 minimal media supplemented with 1% (w/v) glucose and 50 ug/mL Kanamycin.
  • Cells were grown at 37oC with shaking for 12 and 24 hours and OD600 measurements were used to quantitate growth.