# 502: PSEUDOCLOSTRIDIUM MEDIUM


## Main sol. 502

1.2	g	NaCl	
0.4	g	MgCl2 x 6 H2O	
0.3	g	KCl	
0.15	g	CaCl2 x 2 H2O	
0.27	g	NH4Cl	
0.21	g	KH2PO4	
0.1	g	Na2SO4	
1	ml	**Trace element solution SL-10**	
5	g	Inulin	
0.5	ml	Sodium resazurin (0.1% w/v)	
0.03	g	Yeast extract	
0.03	g	Casamino acids (BD Bacto)	
1.5	g	Na2CO3	
2.66	g	Na2HPO4	
1	ml	**Wolin's vitamin solution (10x)**	
0.15	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients (except yeast extract, Casamino acids, vitamins, carbonate, hydrogenphosphate and sulfide), bring medium to the boil, then cool to room temperature under 80% N2 and 20% CO2 gas mixture. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add yeast extract, Casamino acids, hydrogenphosphate and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins are prepared under 100% N2 gas atmosphere and strerilized by filtration. Adjust pH of complete medium to 7.0, if necessay.


## Trace element solution SL-10

10	ml	HCl (25%)	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
990	ml	Distilled water	

1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.


## 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