# 1315: ANAEROBIC THAUERA MEDIUM


## Main sol. 1315

0.85	g	KNO3	
2.52	g	NaHCO3	
0.01	g	CaCl2	
0.5	g	MgSO4 x 7 H2O	
0.01	g	MnSO4	
0.3	g	NH4Cl	
0.05	g	NaCl	
2	ml	**Selenite and tungstate solution**	
884	ml	Distilled water	
100	ml	**Phosphate solution**	
10	ml	**Acetate solution 98.0 g/L**	
5	ml	**Vitamin solution**	
1	ml	**Trace element solution SL-10**	

1. Distribute to gas tight vessels, gas with 90% N2 +10% CO2 to achieve anaerobic conditions and autoclave. After cooling, add the phosphate solution, substrate solution, vitamin solution and trace element solution. After mixing, pH should be 7.1 - 7.5.


## Phosphate solution

1.45	g	Na2HPO4 x 2 H2O	
0.25	g	KH2PO4	
100	ml	Distilled water	

1. Distribute to gas tight vessels under N2, autoclave.


## Acetate solution 98.0 g/L

9.8	g	K-acetate	
100	ml	Distilled water	

1. Filter sterilize and gas with N2. For strains DSM 12141, DSM 12142, DSM 11243 and DSM 12144, acetate may be replaced by Na-capronate (stock solution 2.76 g/100 ml).


## Vitamin solution

50	mg	Vitamin B12	
50	mg	Pantothenic acid	
50	mg	Riboflavin	
10	mg	Pyridoxamine hydrochloride	
20	mg	Biotin	
20	mg	Folic acid	
25	mg	Nicotinic acid	
25	mg	Nicotine amide	
50	mg	alpha-lipoic acid	
50	mg	p-Aminobenzoic acid	
50	mg	Thiamine-HCl x 2 H2O	
1000	ml	Distilled water	

1. Stir for some hours, filter sterilize the 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 and tungstate solution

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

1. First dissolve NaOH, subsequently the metal salts. Distribute to gas tight vessels under N2, autoclave.