ATCC medium: 1754 PETC medium
NH4Cl ....................................1.0 g
KCl......................................0.1 g
MgSo4 . 7H2O .............................0.2 g
NaCl.....................................0.8 g
KH2PO4 ...................................0.1 g
CaCl2 . 2H2O ............................20.0 mg
Yeast extract............................1.0 g
Trace Elements (see below)..............10.0 ml
Wolfe's Vitamin Solution (see below)....10.0 ml
NaHCO3 ...................................2.0 g
Fructose.................................5.0 g
Reducing Agent (see below)..............10.0 ml
Distilled water........................980.0 ml
Add ingredients through vitamins and adjust to pH 5.5. Add NaHCO3, boil, degas and dispense anaerobically under 100% CO2. Autoclave at 121C for 15 minutes. Add fructose and reducing agents from sterile anaerobic stock solutions prepared under 100% N2. Final pH is 5.9.
Trace Elements:
Nitrilotriacetic acid..............2.0 g
MnSO4 . H2O ........................1.0 g
Fe(SO4)2(NH4)2 . 6H2O ...............0.8 g
CoCl2 . 6H2O .......................0.2 g
ZnSO4 . 7H2O .......................0.2 mg
CuCl2 . 2H2O ......................20.0 mg
NiCl2 . 6H2O ......................20.0 mg
Na2MoO4 . 2H2O.....................20.0 mg
Na2SeO4 ...........................20.0 mg
Na2WO4 ............................20.0 mg
Distilled water....................1.0 L
Add nitrilotriacetic acid to water and adjust to pH 6.0 with KOH. Add remainder of ingredients.