Thioglycolate agar is used for the cultivation of aerobic and anaerobic microorganisms in sterility tests and to determine the oxygen requirements of the microorganisms. It is the most frequently used medium in diagnostic bacteriology. When an anaerobic infection is suspected, thioglycollate medium is recommended to isolate strict anaerobes from the blood. This medium does not require an anaerobic container or a special seal for the cultivation of anaerobes. The sodium thioglycolate in the medium consumes oxygen and allows the growth of obligate anaerobes. Thioglycollate agar is also recommended for the cultivation of Clostridium species and for the cultivation of Desulfotomaculum nigrificans.

The enzymatic casein hydrolysate, yeast extract provides nitrogen compounds, vitamin B and other nutrients essential for growth. Dextrose is the fermentable carbohydrate and energy source. Resazurin is the redox indicator. Thioglycollate neutralizes the bacteriostatic effect of mercurial compounds used as preservatives in the injection solution.