# 769: CLOSTRIDIUM VINCENTII MEDIUM


## Main sol. 769

12	g	Sea Salt (SIGMA)	
0.1	g	NH4NO3	
1	g	Yeast extract (BD Bacto)	
0.4	g	Trypticase peptone (BD BBL)	
0.5	ml	Sodium resazurin (0.1% w/v)	
1	g	NaHCO3	
20	ml	Na-phosphate buffer (0.2 M, pH 6.5)	
2	g	Lactose	
1	ml	**Wolin's vitamin solution (10x)**	
0.3	g	L-Cysteine HCl x H2O	
0.3	g	Na2S x 9 H2O	
980	ml	Distilled water	

1. Dissolve ingredients (except bicarbonate, phosphate-buffer, lactose, vitamins, cysteine and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add phosphate-buffer, lactose, cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins are prepared under N2 gas atmosphere and sterilized by filtration. Adjust pH of complete medium to 6.5, if necessary.
2. Note: Sea salts can be replaced by 300.0 ml/l of filtered aged sea water.


## Wolin's vitamin solution (10x)

20	mg	Biotin	
20	mg	Folic acid	
100	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
50	mg	Riboflavin	
50	mg	Nicotinic acid	
50	mg	Calcium D-(+)-pantothenate	
1	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
50	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water