Team:UCLA/Notebook/Recombinant Expression/19 May 2015

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• 1L Expression of Tamura construct
  • Inoculated 950 mL of LB supplemented with 1mL of 100x Carbenicillin (final concentration 100ug/uL) with 10mL starter culture grown overnight.
    • Shook cells in 37 degrees Celsius until an OD600 of 0.6 is reached.
    • Induced protein expression using 5mL of 100mM IPTG stock (final concentration 0.5mM).

• Gene expression monitoring
  • Removed 1mL of culture before adding IPTG and at 1-hour intervals after addition of IPTG, until after 5 hours after gene expression induction.
  • Spun down cells for 15 minutes at 5,300 ref at 4 degrees Celsius, and decanted supernatant.
  • Weighed wet pellet using analytical balance.