Medium data

Search for medium no. = [285].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

285   METHANOSAETA THERMOPHILA MEDIUM

NH4Cl  0.5     g
K2HPO4  0.4     g
MgCl2·6H2O  0.1     g
Resazurin  1.0     mg
Trace minerals (see Medium No. 151)  10.0     ml
Distilled water  1.0     L
Mix ingredients and bring to a boil for 5 min, then cool down under a stream of N2-CO2 (80:20, v/v). Continue the gassing until pH of medium is dropped to 5.8, dispense the medium into culture vessels (e.g., 50 ml in 120 ml serum bottles) under the same gas mixture and seal with butyl rubber stoppers. After autoclaving, add the following anaerobic and sterile stock solutions to the medium (per liter):
5% NaHCO3 solution  20.0     ml
1% CaCl2·2H2O solution  10.0     ml
33% Sodium acetate solution  10.0     ml
Trace vitamins (see Medium No. 197)  10.0     ml
1.42% Coenzyme M solution  10.0     ml
5% Na2S·9H2O solution  5.0     ml
Finally, add filter-sterilized CO2 gas to give 30% (v/v) CO2 in the gas phase. Check the final pH to be 6.5.