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.