ATCC medium: 1803 FYCS medium
(NH4)2SO4.................................1.0 g
KH2PO4 ...................................1.0 g
MgCl2 . 6H2O .............................0.2 g
NaCl.....................................0.2 g
CaCl2 . 2H2O ............................45.0 mg
Wolfe's Mineral Solution (see below)....10.0 ml
Fructose (filter-sterilized).............5.0 g
NaHCO3 (filter-sterilized) ...............1.68 g
Distilled water to.......................1.0 L
Add all ingredients except fructose and sodium bicarbonate.
Autoclave at 121C for 15 minutes. Cool to room temperature and aseptically add fructose and sodium bicarbonate to the concentration indicated from filter-sterilized stock solutions.
Adjust medium for final pH 7.2. Aseptically fill containers to full capacity with sterile medium.
Wolfe's Mineral Solution:
Available from ATCC as a sterile ready-to-use liquid (Trace Mineral Supplement, catalog no. MD-TMS.)
Nitrilotriacetic acid........1.5 g
MgSO4 . 7H2O .................3.0 g
MnSO4 . H2O ..................0.5 g
NaCl.........................1.0 g
FeSO4 . 7H2O .................0.1 g
CoCl2 . 6H2O .................0.1 g
CaCl2 ........................0.1 g
ZnSO4 . 7H2O .................0.1 g
CuSO4 . 5H2O .................0.01 g
AlK(SO4)2 . 12H2O.............0.01 g
H3BO3 ........................0.01 g
Na2MoO4 . 2H2O................0.01 g
Distilled water..............1.0 L
Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound.
Bring volume to 1.0 L with remaining water and add remaining compounds one at a time.