Medium data

Search for medium no. = [231].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

231   METHANOBACTERIUM THERMOAUTOTROPHICUM MEDIUM

KH2PO4  0.3     g
(NH4)2SO4  1.5     g
NaCl  0.6     g
MgSO4·7H2O  0.12    g
CaCl2·2H2O  0.08    g
FeSO4·7H2O  4.0     mg
K2HPO4  0.15    g
Na2CO3  4.0     g
Trace vitamins (see Medium No. 197)  10.0     ml
Trace element solution (see below)  10.0     ml
L-Cysteine·HCl·H2O  0.5     g
Na2S·9H2O  0.5     g
Resazurin  1.0     mg
Distilled water  980.0     ml
Mix ingredients, except cysteine·HCl and Na2S·9H2O, and adjust pH to 7.2. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20). Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Separately autoclave cysteine·HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the cysteine·HCl and Na2S·9H2O solutions. Pressurize the inoculated bottles to 100 kPa H2-CO2 (80:20).
Trace element solution:
EDTA·2Na  0.64    g
MgSO4·7H2O  6.2     g
MnSO4·xH2O  0.55    g
NaCl  1.0     g
FeSO4·7H2O  0.1     g
CoCl2·6H2O  0.17    g
CaCl2·2H2O  0.13    g
ZnSO4·7H2O  0.18    g
CuSO4·5H2O  0.05    g
KAl(SO4)2·12H2O  0.018   g
H3BO3  0.01    g
Na2MoO4·2H2O  0.011   g
NiCl2·6H2O  0.025   g
Dissolve EDTA·2Na first in 500 ml of distilled water. Then add the remaining salts and adjust volume to 1000 ml.