Main sol. 902

Equipment needed: Autoclave Hungate tubes Filter Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NaCl	20.00	g	19.96	341.548
KCl	0.70	g	0.699	9.371
MgSO₄ x 7 H₂O	3.90	g	3.892	15.792
CaCl₂ x 2 H₂O	0.40	g	0.399	2.715
NH₄Cl	0.30	g	0.299	5.597
Na₂HPO₄	0.15	g	0.15	1.055
Na₂SiO₃	0.03	g	0.03	0.245
Yeast extract	0.05	g	0.05	-
Trace element solution SL-10	1.00	ml	-	-
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	0.002
NaHCO₃	0.50	g	0.499	5.94
Na-pyruvate	2.50	g	2.495	22.673
Wolin's vitamin solution (10x)	1.00	ml	-	-
L-Cysteine HCl x H₂O	0.45	g	0.449	2.557
Na₂S x 9 H₂O	0.45	g	0.449	1.87
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients except bicarbonate, pyruvate, vitamins and reducing agents, then adjust pH of solution to 7.0 with NaOH. Sparge medium with 100% N₂ gas for 30 - 45 min to make it anoxic, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add pyruvate, vitamins, cysteine 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 should be sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary.

902	CARCBOXYDIBRACHIUM MEDIUM