KH2PO4 ...................................0.8 g
Sodium acetate...........................2.0 g
Na2SO4 ..................................10.0 g
NaCl....................................30.0 g
1 M Sodium carbonate buffer, pH 9.0....200.0 ml
SLA Trace Elements (see below)...........1.0 ml
VA Vitamins (see below)..................1.0 ml
Distilled water........................600.0 ml
Dissolve ingredients in water. Bring volume to 975 ml and autoclave.
Cool and add aseptically the following sterile solutions:
1% MgCl2 . 6H2O .........................10.0 ml
1% CaCl2 . 2H2O ..........................5.0 ml
20% NH4Cl ................................4.0 ml
5% Na2S . 9H2O (freshly prepared) ........5.0-10.0 ml
Distribute medium immediately into culture bottles.
Allow medium to incubate 2 days prior to inoculation.
SLA Trace Elements: CoCl2 . 6H2O ...........................250.0 mg
NiCl2 . 6H2O ............................10.0 mg
CuCl2 . 2H2O ............................10.0 mg
MnCl2 . 4H2O ............................70.0 mg
ZnCl2 ..................................100.0 mg
H3BO3 ..................................500.0 mg
Na2MoO4 . 2H2O...........................30.0 mg
Na2SeO3 . 5H2O...........................10.0 mg
FeCl2 . 4H2O .............................1.8 g
Distilled water..........................1.0 L
Adjust trace element solution to pH 2-3.
VA Vitamins:
Biotin..................................50.0 mg
Nicotinamide...........................175.0 mg
Thiamine . HCl.........................150.0 mg
p-Aminobenzoic acid....................100.0 mg
Pyridoxine . HCl........................50.0 mg
Calcium D-(+)-pantothenate..............50.0 mg
Cyanocobalamin..........................25.0 mg
Distilled water........................500.0 ml