Main sol. 1301

Equipment needed: Autoclave Hungate tubes Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
KH₂PO₄	0.41	g	0.41	3.013
Na₂HPO₄	0.43	g	0.43	3.029
MgCl₂ x 6 H₂O	0.10	g	0.1	0.492
CaCl₂ x 2 H₂O	0.11	g	0.11	0.748
NaCl	0.30	g	0.3	5.133
NH₄Cl	0.30	g	0.3	5.609
Trace element solution	1.00	ml	-	-
Selenite-tungstate solution	1.00	ml	-	-
Yeast extract	0.20	g	0.2	-
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	-
Na₂CO₃	1.50	g	1.5	14.153
D-Glucose	1.80	g	1.8	9.991
Wolin's vitamin solution	2.00	ml	-	-
Na₂S x 9 H₂O	0.30	g	0.3	1.249
L-Cysteine HCl x H₂O	0.50	g	0.5	2.847
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients (except carbonate, glucose, vitamins and reducing agents) and 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, vitamins, sulfide and cysteine from sterile anoxic stock solutions prepared under 100% N₂ gas and carbonate from a sterile stock solution prepared under 80% N₂ and 20% CO₂ gas mixture. Adjust pH of the complete medium to 7.0-7.2, if necessary.

1301	TEPIDANAEROBACTER ACETATOXYDANS MEDIUM