Main sol. 1062

Equipment needed: Autoclave Hungate tubes Filter Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
KH₂PO₄	0.20	g	0.198	1.454
NH₄Cl	0.25	g	0.247	4.623
NaCl	1.00	g	0.989	16.925
MgCl₂ x 6 H₂O	0.40	g	0.396	1.946
KCl	0.50	g	0.495	6.634
CaCl₂ x 2 H₂O	0.15	g	0.148	1.009
Modified Wolin's mineral solution	10.00	ml	-	-
Sodium resazurin (0.1% w/v)	0.50	ml	4.9e-4	0.002
Na₂CO₃	1.50	g	1.484	13.999
FeSO₄ x 7 H₂O	22.00	mg	0.022	0.078
Seven vitamins solution	1.00	ml	-	-
Yeast extract	2.00	g	1.978	-
Na-pyruvate	4.50	g	4.451	40.448
Na₂-fumarate	6.50	g	6.429	40.174
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients (except carbonate, ferrous iron sulfate, vitamins and organic substrates), 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 ferrous iron sulfate (dissolved in 0.1 N H₂SO₄), vitamins, pyruvate, fumarate and yeast extract 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 atmosphere. Stock solutions of ferrous iron sulfate, vitamins, pyruvate and fumarate should be sterilized by filtration. Adjust pH of the complete medium to 7.5, if necessary.
2 Note: Addition of 10 - 20 mg sodium dithionite per litre (e.g. from 5% (w/v) solution, freshly prepared under N₂ and filter-sterilized) may stimulate growth at the beginning.

1062	DESULFITOBACTERIUM (PCE II) MEDIUM