Main sol. 899

Equipment needed: Hungate tubes Filter Autoclave Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NH₄Cl	1.00	g	0.99	18.51
K₂HPO₄	0.30	g	0.297	1.705
KH₂PO₄	0.30	g	0.297	2.183
MgCl₂ x 6 H₂O	1.00	g	0.99	4.87
CaCl₂ x 2 H₂O	0.10	g	0.099	0.673
KCl	0.20	g	0.198	2.656
NaCl	0.60	g	0.594	10.165
Modified Wolin's mineral solution	10.00	ml	-	-
Yeast extract	2.00	g	1.98	-
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	0.002
Na₂CO₃	1.00	g	0.99	9.342
D-Glucose	4.00	g	3.96	21.983
Na₂S₂O₃ x 5 H₂O	2.50	g	2.475	9.973
L-Cysteine HCl x H₂O	0.50	g	0.495	2.819
Na₂S x 9 H₂O	0.45	g	0.446	1.855
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients (except carbonate, glucose, thiosulfate, cysteine 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 and autoclave. Add glucose, thiosulfate (sterilized by filtration), cysteine 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. The pH of the complete medium should be at 7.0.

899	CALDANAEROBACTER MEDIUM