# 14: CLOSTRIDIUM FORMICACETICUM MEDIUM


## Main sol. 14

5	g	Yeast extract	
10	g	K2HPO4	
10	ml	**Trace element solution SL-4**	
1	ml	Pyridoxine hydrochloride (0.1% w/v)	
0.75	g	Na-thioglycolate	
0.5	ml	Sodium resazurin (0.1% w/v)	
1	g	Na2CO3	
5	g	D-Fructose	
15	g	Agar, for solid medium (optional)	
1000	ml	Distilled water	

1. Dissolve ingredients (except carbonate and fructose) and sparge medium with 100% N2 gas for at least 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add fructose from a sterile anoxic stock solution prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Adjust pH of complete medium to 8.0.


## 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.