Main sol. 292

Equipment needed: Filter Hungate tubes Gassing station Autoclave

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Glycine	1.50	g	1.496	19.922
Na₂SO₄	3.00	g	2.991	21.058
KH₂PO₄	0.20	g	0.199	1.465
MgCl₂ x 6 H₂O	0.40	g	0.399	1.962
NaCl	1.20	g	1.196	20.473
CaCl₂ x 2 H₂O	0.15	g	0.15	1.017
KCl	0.40	g	0.399	5.349
Yeast extract	0.20	g	0.199	-
Trace element solution SL-10	1.00	ml	-	-
Selenite-tungstate solution	1.00	ml	-	-
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	0.002
NaHCO₃	2.00	g	1.994	23.736
Wolin's vitamin solution (10x)	1.00	ml	-	-
Na₂S x 9 H₂O	0.30	g	0.299	1.245
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients (except bicarbonate, vitamins and sulfide), adjust pH to 6.5, then sparge with 100% N₂ gas for 30 - 45 min to make it anoxic and dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials. After autoclaving complete the medium by adding vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas mixture. Vitamins are sterilized by filtration. Adjust pH of the complete medium to 7.4 - 7.5, if necessary.

292	ACIDAMINOBACTER MEDIUM