Medium data

Search for medium no. = [195].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

195 THERMOPROTEUS NEUTROPHILUS MEDIUM

Modified Brock's salt base solution (see Medium No. 165)	`1.0`	L
NaHCO₃	`0.85`	g
Yeast extract (BD-Difco)	`0.2`	g
Sodium dithionite	`25.0`	mg
Sulfur (powder)	`8.0`	g
Resazurin	`0.4`	mg

Mix ingredients, except yeast extract, sodium dithionite and sulfur, and adjust pH to 6.5. Autoclave, then cool under H₂-CO₂ (80:20) gas mixture. Separately autoclave yeast extract as a 10% solution under a N₂ atmosphere. Filter-sterilize NaHCO₃ as an 8% solution while gassing with N₂. Steam sulfur for 3 hr on each of 3 successive days. Aseptically and anaerobically add the yeast extract and NaHCO₃ solutions, and sulfur to the medium under a H₂-CO₂ (80:20) atmosphere. Readjust pH to 6.5 by further gassing with H₂-CO₂ (80:20) gas mixture, if necessary. Prior to inoculation, add filter-sterilized 2.5% sodium dithionite solution. Pressurize inoculated bottles to 100 kPa H₂-CO₂ (80:20).