Main sol. 827

Equipment needed: Hungate tubes Filter Gassing station Autoclave Water bath

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NH₄Cl	0.33	g	0.329	6.157
KCl	0.33	g	0.329	4.418
KH₂PO₄	0.33	g	0.329	2.42
MgCl₂ x 6 H₂O	0.33	g	0.329	1.62
CaCl₂ x 2 H₂O	0.33	g	0.329	2.24
Trace element solution SL-10	1.00	ml	-	-
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	0.002
Sulfur (powdered)	10.00	g	9.98	311.195
Na₂CO₃	1.00	g	0.998	9.416
Yeast extract	0.50	g	0.499	-
D-Glucose	5.00	g	4.99	27.698
Wolin's vitamin solution (10x)	1.00	ml	-	-
Na₂S x 9 H₂O	0.50	g	0.499	2.078
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients except sulfur, carbonate, yeast extract, glucose, vitamins, and sulfide, then sparge medium with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur. Heat the medium vials to 90-100°C for 1 hour on each of 3 successive days to sterilize the medium. Add yeast extract, glucose, vitamins (sterilized by filtration), and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas mixture. Prior to use check pH of complete medium and adjust to 6.8 - 7.0, if necessary.

827	THERMOANAEROBACTER SULFUROPHILUS MEDIUM