IGEM:Virginia 2012/Protocols/Preparing a Pertussis Culture

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(New page: ==Overview== Below is the procedure for preparing a ''Bordetella pertussis'' culture from frozen stock. First, the bacteria will be grown on a plate for ~3 days. Then it will be grown in ...)
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==Overview==
==Overview==
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Below is the procedure for preparing a ''Bordetella pertussis'' culture from frozen stock. First, the bacteria will be grown on a plate for ~3 days. Then it will be grown in liquid culture for a day, and then diluted so that the bacteria will be in log phase a day later. This procedure should be done in a hood to prevent aerosolization.
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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
==Materials==
==Materials==
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*Pertussis frozen stock
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*Pertussis frozen stock (keep on ice)
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*Sheep's blood agar plates
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*Sheep blood agar plates
*Sterilized sticks
*Sterilized sticks
==Procedure==
==Procedure==
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#  
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Note: This procedure should be done in a hood to prevent aerosolization.
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# Using a sterilized stick, streak the entire surface of the plate. Discard the stick in the biohazard container.
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# Put the plates in a plastic bag to prevent drying out and incubate at 37°C for three days.
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# Inoculate the liquid culture. Incubate at 35.5°C.
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# The next day, dilute the liquid culture.
==Notes==
==Notes==

Revision as of 13:26, 31 May 2012

Contents

Overview

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

Materials

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

Procedure

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.

Notes

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


References

Contact

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