# 1250: DESULFOSPOROSINUS ACIDOPHILUS MEDIUM


## Main sol. 1250

0.45	g	(NH4)2SO4	
0.5	g	MgSO4 x 7 H2O	
0.05	g	KCl	
0.02	g	Ca(NO3)2 x 4 H2O	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
0.5	g	Yeast extract	
0.5	ml	Sodium resazurin (0.1% w/v)	
10	g	KH2PO4	
1	g	Na2HPO4	
3.5	g	D-Fructose	
1	ml	**Wolin's vitamin solution (10x)**	
0.5	g	L-Cysteine HCl x H2O	
1000	ml	Distilled water	

1. Dissolve ingredients (except phosphates, vitamins, fructose and cysteine) and 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. Prepare a phosphate buffer stock solution with KH2PO4 (25% w/v) and Na2HPO4, adjust pH of the buffer to 5.8. Prior to inoculation add phosphate buffer, vitamins, fructose and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins and D-fructose should be sterilized by filtration. The pH of the complete medium should be at 5.8.


## 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.


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled 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