# 211: THERMOANAEROBACTERIUM MEDIUM


## Main sol. 211

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

1. Dissolve ingredients (except glucose, vitamins and sulfide), adjust pH to 6.0, sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense under same gas atmosphere into Hungate-type tubes or serum vials and autoclave. After sterilization add glucose and sulfide from anoxic stock solutions autoclaved under 100% N2 gas and vitamins from a filter-sterilized anoxic stock solution prepared under 100% N2. Adjust pH of complete medium to 6.0 - 6.5, if necessary.


## 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