Medium data

Search for medium no. = [945].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

945   MB MEDIUM FOR METHANOBACTERIUM

KH2PO4  0.15    g
K2HPO4  0.3     g
NaCl  0.6     g
(NH4)2SO4  0.3     g
MgSO4·7H2O  0.12    g
CaCl2·2H2O  0.08    g
Trace minerals solution (see below)  10.0     ml
Resazurin  1.0     mg
Distilled water  932.0     ml
Mix components thoroughly, and autoclave under a N2-CO2 (80:20, v/v) gas atmosphere. After cooling, aseptically and anaerobically add the following solutions (filter-sterilized):
8% NaHCO3 solution  48.0     ml
Trace vitamins (see Medium No. 197)  10.0     ml
Distribute the medium into culture vessels (leave >80% gas phase by volume) under a H2-CO2 (80:20, v/v) gas mixture, seal with butyl rubber stoppers. Finally, add per liter the following solutions from anaerobic stocks (autoclaved and stored under a N2 atmosphere):
5% L-Cysteine·HCl·H2O solution  6.0     ml
5% Na2S·9H2O solution  6.0     ml
Pressurize the inoculated vessels to 200 kPa H2-CO2 (80:20, v/v).
Trace minerals solution:
Nitrilotriacetic acid  1.5     g
MgSO4·7H2O  3.0     g
MnSO4·xH2O  0.5     g
FeSO4·7H2O  0.1     g
CoSO4·7H2O  0.1     g
ZnSO4·7H2O  0.1     g
CuSO4·5H2O  0.01    g
H3BO3  0.01    g
Na2MoO4·2H2O  0.01    g
Distilled water  1.0     L
Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. Adjust final pH to 7.0.