# 507a: CARBOXYDOTHERMUS SIDEROPHILUS MEDIUM


## Main sol. 507a

0.33	g	KCl	
0.52	g	MgCl2 x 6 H2O	
0.29	g	CaCl2 x 2 H2O	
0.33	g	NH4Cl	
0.33	g	KH2PO4	
8.25	g	Na2-9,10-anthraquinone-2,6-disulfonate (TCI A0308)	
10	ml	**Trace element solution SL-4**	
1	g	NaHCO3	
0.2	g	Yeast extract	
1	ml	**Wolin's vitamin solution (10x)**	
1000	ml	Distilled water	

1. Dissolve ingredients except bicarbonate, yeast extract, vitamins and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. After completing the medium adjust pH to 6.8 - 7.0.
2. Inoculated vessels are pressurized with sterile carbon monoxide gas to 1 bar overpressure.


## Trace element solution SL-4

0.5	g	Na2-EDTA	
0.2	g	FeSO4 x 7 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 2 H2O	
0.02	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds.


## Wolin's vitamin solution (10x)

20	mg	Biotin	
20	mg	Folic acid	
100	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
50	mg	Riboflavin	
50	mg	Nicotinic acid	
50	mg	Calcium D-(+)-pantothenate	
1	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
50	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water