# 1661: ALKALIPHILUS NAMSARAEVII MEDIUM


## Main sol. 1661

0.5	g	KH2PO4	
25	g	NaCl	
2.5	g	Yeast extract	
1.85	g	NaHCO3	
1.2	g	Na2CO3	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
7.3	g	Fe(III) citrate (19% Fe)	
0.2	g	MgCl2 x 6 H2O	
1	ml	**Wolin's vitamin solution (10x)**	
1000	ml	Distilled water	

1. Dissolve ingredients (except bicarbonate, carbonate, trace elements, ferric citrate, magnesium chloride and vitamins) and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate and carbonate, then adjust pH to 9.5 and dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add ferric citrate (10% w/v stock solution; dissolve by boiling and adjust pH to 7.5-8.0), trace elements, selenite-tungstate solution, magnesium chloride and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas. Sterilize vitamins by filtration. Adjust pH of the complete medium to 9.5, if necessary.


## 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