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).