ATCC medium: 1552 Modified Van Niel's yeast medium with pyruvate
MgSO4 ..............................0.1 g
EDTA...............................2.0 mg
Yeast extract.....................10.0 g
Distilled water..................885.0 ml
Adjust medium for final pH 7.1, +/- 0.1. Autoclave at 121C for 15 minutes.
Cool to room temperature and aseptically add the following filtersterilized solutions:
Trace Metal Mix A-5 (see below)....1.0 ml
Trace Metals B (see below)........10.0 ml
4% K2HPO4 ..........................5.0 ml
1.1% Sodium pyruvate.............100.0 ml
Trace Metal Mix A5: H3BO3 ..............................2.86 g
MnCl2 . 4H2O .......................1.81 g
ZnSO4 . 7H2O .......................0.222 g
Na2MoO4 . 2H2O......................0.39 g
CuSO4 . 5H2O .......................0.079 g
Co(NO3)2 . 6H2O....................49.4 mg
Distilled water....................1.0 L
Trace Metals B:
Ferric ammonium citrate............0.2 g
CaCl2 . 2H2O .......................0.3 g
Distilled water..................100.0 ml