IGEM:Virginia 2012/Protocols/Preparing a Pertussis Culture: Difference between revisions
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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== | ||
Below is the procedure for preparing a ''Bordetella pertussis'' culture | 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== | ||
*Pertussis frozen stock | *Pertussis frozen stock (keep on ice) | ||
*Sheep | *Sheep blood agar plates | ||
*Sterilized sticks | *Sterilized sticks | ||
==Procedure== | ==Procedure== | ||
# | Note: This procedure should be done in a hood to prevent aerosolization. | ||
# Using a sterilized stick, streak the entire surface of the plate. Discard the stick in the biohazard container. | |||
# Put the plates in a plastic bag to prevent drying out and incubate at 37°C for three days. | |||
# Inoculate the liquid culture. Incubate at 35.5°C. | |||
# The next day, dilute the liquid culture. | |||
==Notes== | ==Notes== | ||
Revision as of 11:26, 31 May 2012
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.
- Using a sterilized stick, streak the entire surface of the plate. Discard the stick in the biohazard container.
- Put the plates in a plastic bag to prevent drying out and incubate at 37°C for three days.
- Inoculate the liquid culture. Incubate at 35.5°C.
- 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
- Who has experience with this protocol?
or instead, discuss this protocol.
