# 143: CLOSTRIDIUM SP. LA1 MEDIUM


## Main sol. 143

6	g	Crotonic acid	
0.3	g	NaOH	
0.15	g	(NH4)2HPO4	
0.1	g	K2HPO4	
0.03	g	MgCl2 x 6 H2O	
0.04	g	CaCl2 x 2 H2O	
0.05	g	NH4Cl	
0.6	ml	MgSO4 x 7 H2O (0.1% w/v)	
0.4	ml	MnSO4 x 2 H2O (0.1% w/v)	
0.4	ml	FeSO4 x 7 H2O (0.1% w/v in 0.1 N H2SO4)	
10	mg	(NH4)6Mo7O24 x 4 H2O	
0.04	ml	Biotin (0.1% w/v)	
0.8	ml	p-Aminobenzoic acid (0.1% w/v)	
1	g	Tryptone (BD BBL)	
1	g	Yeast extract (OXOID)	
0.5	ml	Sodium resazurin (0.1% w/v)	
4	g	K2CO3	
1000	ml	Distilled water	

1. Dissolve ingredients (except potassium carbonate) 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 and autoclave. An anoxic stock solution of the potassium carbonate is autoclaved separately under 80% N2 and 20% CO2 gas mixture and thereafter added to the sterile medium. Adjust pH of the complete medium to 6.8 - 7.0. Prior to inoculation add 10 - 20 mg/l sodium dithionite from a 5% (w/v) stock solution that has been freshly prepared under 100% N2 gas and sterilized by filtration to reduce the medium.