# 1687: THERMOANAEROBACULUM MEDIUM


## Main sol. 1687

994	ml	**Solution A**	
0.1	ml	**Solution B**	
0.1	ml	**Solution C**	

1. Sparge solution A with 100% N2 gas for 30 - 45 min to make it anoxic, distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solutions B and C are autoclaved separately under 100% N2 gas. Adjust pH of complete medium to 7.2 - 7.4, if necessary.
2. Use 1.10 g/l Na-pyruvate as substrate. Na-pyruvate is added to the autoclaved medium from a sterile anoxic stock solution sterilized by filtration.


## Solution A

0.02	g	KH2PO4	
0.025	g	NH4Cl	
0.1	g	NaCl	
0.04	g	MgCl2 x 6 H2O	
0.05	g	KCl	
0.015	g	CaCl2 x 2 H2O	
0.85	g	NaNO3	
0.1	g	Yeast extract	
0.05	ml	Sodium resazurin (0.1% w/v)	
994	ml	Distilled water	


## Solution B

1	ml	**Trace element solution SL-10**	


## Solution C

1	ml	**Selenite-tungstate solution**	


## Trace element solution SL-10

10	ml	HCl (25%)	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
990	ml	Distilled water	

1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water