Procedures for the Determination of Oxygen Relationships:
Anaerobic Jar (Gas Pak)
Brewers Jar, Gas Pak, Palladium catalyst, oxygen indicator strip, 10 ml
1. Inoculate a plate of ENA with the organism to be tested.
2. Place the plate in the Brewers jar, open the corner
of the gas pak and place inside the jar.
3. Fill gas pak with 10 ml DI water, and quickly
apply the indicator strip to the inside of the brewers jar.
4. Seal the lid tightly with your fingers.
Incubate at the appropriate temperture.
5. Only organisms capable of anaerobic growth should grow on
See the setup of a Anaerobic Jar (not yet available)
Thyioglycolate tube with methylene blue indicator and organism to be tested.
1. Inoculate a thioglycolate tube with the organism to
be tested for oxygen relationship
2. Grow at optimal growth temperature
3. Observe growth patterns
Results of thyioglycolate broth (not yet available)
ENA slant w/ cotton plug, cork stopper, 4% NaOH solution, Pyrogallic acid
1. Inoculate the ENA slant with the organism to be tested.
2. Push the cotton plug down into the tube with a
sterile loop until it sits directly above the slant.
3. Add pyrogallic acid crystals to the space above
4. Add approximately 2 ml of the NaOH solution and
cork the slant.
5. Incubate inverted at optimal temperature to avoid excess
NaOH dropping onto the growing culture.
See a Buchner Tube (not yet available)
Boiled ENA allowed to cool to approximately 50 degrees C,
1. Stir a loopful of culture into melted agar by
twisting the loop as you remove it from the bottom to the top of the tube.
2. Replace cap and observe for any areas of visible
3. Allow to solidify standing upright, and incubate
at optimal growth temperature
4. Observe growth patterns
Results of Deep Tubes (not yet available)
Mineral Oil-Sealed Tubes
Boiled ENB allowed to cool to approximately 50 degrees C, dropper of
1. Using a broth culture, inoculate the boiled ENB
without excessively disturbing the media.
2. Pour a layer of mineral oil about 1 cm thick on
the top of the broth
3. Inoculate at optimum temperature and observe growth
Results of Mineal Oil-Sealed Tubes (not yet available)