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).