Main sol. 124

Equipment needed: Hungate tubes Filter Autoclave Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NaCl	1.17	g	1.167	19.961
MgCl₂ x 6 H₂O	0.40	g	0.399	1.962
KCl	0.30	g	0.299	4.012
CaCl₂ x 2 H₂O	0.15	g	0.15	1.017
NH₄Cl	0.27	g	0.269	5.033
KH₂PO₄	0.20	g	0.199	1.465
Na₂SO₄	2.84	g	2.832	19.935
Trace element solution SL-10	1.00	ml	-	-
Selenite-tungstate solution	1.00	ml	-	-
Yeast extract	1.00	g	0.997	-
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	0.002
NaHCO₃	4.50	g	4.487	53.407
Na-acetate	1.40	g	1.396	17.015
Na-butyrate	1.40	g	1.396	12.679
Wolin's vitamin solution (10x)	1.00	ml	-	-
Na₂S x 9 H₂O	0.40	g	0.399	1.66
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients (except bicarbonate, acetate, butyrate, vitamins, and sulfide) and sparge medium with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, adjust pH to 7.0, then dispense medium under 80% N₂ and 20% CO₂ gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add acetate, butyrate, vitamins, and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas. The vitamin solution should be sterilized by filtration. Before use, check the complete medium's pH and adjust it to 7.0 - 7.2, if necessary.

124	DESULFOFARCIMEN MEDIUM