Procedures for the Determination of Oxygen Relationships:


Anaerobic Jar (Gas Pak)

Materials:

Brewers Jar, Gas Pak, Palladium catalyst, oxygen indicator strip, 10 ml pipet.

Procedure:

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 the plate.

See the setup of a Anaerobic Jar (not yet available)
Lab Procedures


Thioglycolate Tube

Materials:

Thyioglycolate tube with methylene blue indicator and organism to be tested.
 

Procedure:

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)
Lab Procedures


Buchner Tube

Materials:

ENA slant w/ cotton plug, cork stopper, 4% NaOH solution, Pyrogallic acid crystals

Procedure:

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 the cotton.
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)
Lab Procedures



Deep Tube

Materials:

Boiled ENA allowed to cool to approximately 50 degrees C,

Procedure:
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 inoculum
3.    Allow to solidify standing upright, and incubate at optimal growth temperature
4.    Observe growth patterns

Results of Deep Tubes (not yet available)
Lab Procedures



Mineral Oil-Sealed Tubes

Materials:

Boiled ENB allowed to cool to approximately 50 degrees C, dropper of mineral oil

Procedure:
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 patterns

Results of Mineal Oil-Sealed Tubes (not yet available)
Lab Procedures