# 54b: CLOSTRIDIUM SP. MEDIUM


## Main sol. 54b

20	g	Glucose	
10	g	Yeast extract	
20	g	CaCO3 (light precipitate)	
17	g	Agar, for solid medium (optional)	
1000	ml	Distilled water	

1. Dissolve ingredients (note that most of the carbonate remains suspended) and sparge medium with 100% N2 gas for 30 -45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials vials while stirring to keep carbonate suspended and autoclave.