ATCC medium: 1782 Eubacterium angustum medium
Trizma Base (Sigma T 1503)...............3.0 g
Uric acid................................3.0 g
NH4Cl ....................................0.5 g
MgSO4 . 7H2O .............................0.05 g
K2HPO4 ...................................0.1 g
Wolfe's Mineral Solution (see below)....10.0 ml
Selenium Solution (see below)............1.0 ml
Yeast extract............................1.0 g
Resazurin................................1.0 mg
L-Cysteine . HCl.........................0.5 g
Na2S . 9H2O .............................50.0 mg
NaHCO3 ...................................5.0 g
Distilled water..........................1.0 L
Dissolve uric acid in distilled water and Trizma with heat and stirring. Add remaining ingredients except reducing agents and sodium bicarbonate. Heat to boiling and cool under 80% N2, 10% CO2, 10% H2. Add bicarbonate and reducing agents. Tube anaerobically under same gas phase. Autoclave at 121C for 15 minutes. Final pH 7.9 +/- 0.1. 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.
Selenium Solution: Na2SeO3 . 5H2O......................3.0 mg
0.01M NaOH.........................1.0 L
Use 1 ml/L of medium.