# 255: CLOSTRIDIUM (GS) MEDIUM


## Main sol. 255

0.5	g	KH2PO4	
1	g	K2HPO4 x 3 H2O	
2	g	Urea	
0.5	g	MgCl2 x 6 H2O	
0.05	g	CaCl2 x 2 H2O	
1.25	ml	FeSO4 x 7 H2O (0.1% w/v in 0.1 N H2SO4)	
10	g	Morpholinopropane sulfonic acid	
0.5	ml	Sodium resazurin (0.1% w/v)	
6	g	Yeast extract	
1	g	L-Cysteine HCl x H2O	
5	g	Cellobiose	
1000	ml	Distilled water	

1. Dissolve ingredients except cysteine and cellobiose and adjust pH to 7.2. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then add cysteine and readjust pH to 7.2, if necessary. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Prior to inoculation add cellobiose from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration.