# 206: THERMODESULFOBACTERIUM MEDIUM


## Main sol. 206

3	g	Na2SO4	
1	g	NH4Cl	
0.2	g	MgCl2 x 6 H2O	
0.3	g	KH2PO4	
2	g	Na2HPO4 x 12 H2O	
10	ml	**Trace element solution**	
1.5	ml	FeSO4 x 7 H2O (0.1% w/v in 0.1 N H2SO4)	
0.5	ml	Sodium resazurin (0.1% w/v)	
1	g	Yeast extract	
4	g	Na-L-lactate	
5	ml	**Wolin's vitamin solution**	
0.5	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients (except yeast extract, lactate, vitamins and sulfide) and sparge medium for 30 - 45 min with 100% N2 gas to make it anoxic. Distribute under same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add yeast extract, lactate, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Before use, neutralize the sodium sulfide solution by dropwise addition of 2 N H2SO4 (see medium 29). Adjust pH of complete medium to 6.8 - 7.0.


## Trace element solution

12.8	g	Nitrilotriacetic acid (NTA)	
0.2	g	FeCl2 x 4 H2O	
0.1	g	MnCl2 x 4 H2O	
0.17	g	CoCl2 x 6 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	ZnCl2	
0.02	g	CuCl2	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
0.03	g	NiCl2 x 6 H2O	
1	g	NaCl	
0.03	g	Na2SeO3 x 5 H2O	
1000	ml	Distilled water	

1. First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.00 ml.


## Wolin's vitamin solution

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium D-(+)-pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water