ATCC Medium: 2722 Sup plemented Trypti c Soy Broth/Agar
Tryptic Soy Broth… ………… …………… …………………..30.0 g
Yeast Extract…… …………… ………………………… ……..5.0 g
L-cysteine hydrochloride…… ………………………… …….0.5 g
Hemin tock (see below)………………… …………..……...1.0 ml
Vitamin K1 Stock* (5 mg/ml)… ………… ……………………0.2 ml
Agar (If required)… ………… ………… ……………………15.0 g
DI Water………… …………… ………………………… ……...100 0 ml
Autocla ve at 121?C . Wrap ea ch bottle in aluminum foil or kee p bottle in the dark as hemin and vitamin K are light sensitive and will be degraded. Suppleme nted TSB can be pre-reduced an d pre-warm ed by incubating a bo ttle of fres h media in the anaerobic chamber overnight before usi ng it. *Men adione (vitamin K3) can be use at a final concentration of 1.0 mcg/ml i stead of vitamin K1.
Hemin Stock Solu tion Hemin ………… …………………….0.5 g
K2HPO4…………… ………… ……..1.74 g
DI Water………… …………… ……….100 ml
Boil the above ingredients in a large flas k. Store in the dark. Note: H emin will h ve a final concentration of 5 mc g/ml. Vita in K1 will have a final concentration of 1.0 mcg/ml Final Pr oduct Des cription ?Medium should be translucent and tan in color. ?Recommende d storage emperature: 2-8?C.