ATCC Medium 2722: Supplemented Tryptic Soy Broth

ATCC Medium 2722: Supplemented Tryptic Soy Broth
Tryptic Soy Broth…………………………………………… 30.0 g
Yeast Extract………………………………………………... 5.0 g
L-cysteine hydrochloride…………………………………… 0.5 g
Hemin Stock (see below) …………………………………. 1.0 mL
Vitamin K1 Stock* (5 mg/ml) ……………………………… 0.2 mL
DI Water……………………………………………………... 1000 mL
Autoclave at 121ºC.

Wrap each bottle in aluminum foil or keep bottle in the dark as hemin and vitamin
K are light sensitive and will be degraded.

Supplemented TSB can be pre-reduced and pre-warmed by incubating a bottle
of fresh media in the anaerobic chamber overnight before using it.

*Menadione
(vitamin K3) can be used at a final concentration of 1.0 mcg/ml instead of vitamin
K1.
Hemin Stock Solution
Hemin …………………………………… 0.5 g
K2HPO4…………………………………. 1.74 g
DI Water………………………………… 100 mL
Boil the above ingredients in a large flask. Store in the dark.
Note: Hemin will have a final concentration of 5 mcg/mL. Vitamin K1 will have a
final concentration of 1.0 mcg/mL
Final Product Description
➢ Medium should be translucent and tan in
color.
➢ Recommended storage temperature: 2-8ºC.