Middlebrook 7H9 Broth………………………………………..4.7 g
Tween 80……………………………………………………….0.5 g
Mycobactin J*…………………………………………………..2.0 mg
Dubos Oleic Albumin Complex (OADC) (see below)……...100.0 ml
DI Water………………………………………………………...900.0 ml
Add and mix all components except for OADC. Autoclave at 121?C, cool to room temperature and aseptically add OADC.
OADC
Oleic Acid (Sigma Cat # O-1008)……………………………..0.05 g
Albumin Fraction V Bovine (Sigma A9647)……….………...5 g
Dextrose (BD 215530)…………………………………………2 g
Catalase (beef) (Sigma C-9322)……………………………...0.004 g
NaCl……………………………………………………………..0.85 g
DI Water…………………………………………………………100 ml
OADC is available from BD (237510) or can be made following the formulation above.
*Note: Mycobactin J is stored dehydrated in 2 mg quantities unless otherwise specified. Rehydrate w/ 2mL ethanol or 1mL/mg. (Available from Allied Laboratories, Inc., 2520 Hunt St. Ames, IA 50010).