IGEM:Virginia 2012/Protocols/Preparing a Pertussis Culture

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Below is the procedure for preparing a Bordetella pertussis culture. First, the frozen stock is streaked onto a sheep blood agar plate. This plate is then incubated for ~3 days, until it produces pertussis colonies which are gray/white and dull rather than shiny. These colonies are then used to inoculate a liquid culture. Since pertussis is aerobic, the liquid culture must have a high amount of surface area. This liquid culture can then be diluted so that it is in the logarithmic growth phase


  • Pertussis frozen stock (keep on ice)
  • Sheep blood agar plates
  • Sterilized sticks


Note: This procedure should be done in a hood to prevent aerosolization.

  1. Using a sterilized stick, streak the entire surface of the plate. Discard the stick in the biohazard container.
  2. Put the plates in a plastic bag to prevent drying out and incubate at 37°C for three days.
  3. Inoculate the liquid culture. Incubate at 35.5°C.
  4. The next day, dilute the liquid culture.


Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!



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