Medium data

Search for medium no. = [524].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

524   METHANE OXIDIZING BACTERIUM MEDIUM

NaCl  30.0     g
KH2PO4  0.14    g
CaCl2·2H2O  0.8     g
MgSO4·7H2O  3.4     g
MgCl2·6H2O  4.2     g
KCl  0.33    g
NaNO3  0.25    g
NH4Cl  0.25    g
Na2SiO3·9H2O  0.2     g
Fe2(SO4)3·xH2O  20.0     mg
Na2SeO3·5H2O  0.5     mg
NiCl2·6H2O  0.5     mg
Trace mineral solution (see Medium No. 523)  3.0     ml
Distilled water  1.0     L
Mix components, adjust pH to 6.0 - 6.5 and autoclave. After cooling, add the following solutions (filter-sterilized) to the medium:
Trace vitamins (see Medium No. 197)  1.0     ml
5% NaHCO3 solution  10.0     ml
1 mM CuSO4 solution  2.0     ml
Distribute the medium into culture vessels (leave >80% volume gas phase) under a N2-CO2 (4:1) gas mixture and seal with butyl rubber stoppers. Pressurize the culture vessels to 200 kPa with methane gas. Finally add O2 gas to make up 2% O2 concentration in the gas phase. Readjust pH to 6.0 - 6.3, if necessary.