Medium data

Search for medium no. = [236].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

236   THERMOCLADIUM MEDIUM

Modified Brock's salt base solution (see Medium No. 165)  1.0     L
Trisodium citrate·2H2O  2.94    g
Yeast extract (BD-Difco)  0.5     g
Trace vitamins (see Medium No. 197)  10.0     ml
Resazurin  1.0     mg
Na2S·9H2O  0.5     g
Sulfur (powder)  10.0     g
Combine Modified Brock's salt base solution, trisodium citrate and resazurin, and adjust pH to 3.5 with 10 N H2SO4. Autoclave under a N2 atmosphere. Separately autoclave yeast extract and Na2S·9H2O as 10% and 5% solutions, respectively, under a N2 atmosphere. Filter-sterilize Trace vitamins. Steam sulfur for 3 hr on each of 3 successive days. Aseptically mix components except Na2S·9H2O in suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) while gassing with H2-CO2 (80:20) gas mixture and seal with butyl rubber stoppers. Prior to inoculation, aseptically and anaerobically add the Na2S·9H2O solutions and adjust pH to 4.0-4.5, if necessary. When the indicator (resazurin) turns to colorless, inoculate the culture and pressurize the vessels to 100 kPa H2-CO2 (80:20).