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== | ||
| - | 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 14:26, 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.
- 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.
