# 590: DICHOTOMICROBIUM THERMOHALOPHILUM MEDIUM


## Main sol. 590

1	g	Yeast extract (Difco, Bacto )	
1	g	Na2-DL-malate	
20	ml	**Hutner's salts**	
980	ml	**Artificial sea water, 3 times concentrated**	
18	g/l	Agar, for solid medium	
28.4	ml	NaHCO3 (8 %)	

1. pH 7.0-7.2
2. Agar at a concentration of 18 g/l is added for solid media.
3. Prepare an 8% solution of NaHCO3 in half full 100ml serum bottles closed with a rubber
septum and autoclave. Add 28.4 ml of the sterile 8% solution of NaHCO3 (2.822g/l) per
litre of the sterile, cooled medium.


## Artificial sea water, 3 times concentrated

70.43	g	NaCl	
11.75	g	Na2SO4	
2.88	g	NaHCO3	
1.99	g	KCl	
0.29	g	KBr	
31.86	g	MgCl2 x 6 H2O	
4.35	g	CaCl2 x 2 H2O	
0.08	g	H3BO3	
1000	ml	Distilled water	

1. Add NaHCO3 from a filter-sterilized stock solution when the medium has cooled.


## Hutner's salts

10	g	Nitrilotriacetic acid (NTA)	
29.7	g	MgSO4 x 7 H2O	
3.335	g	CaCl2 x 2 H2O	
9.25	mg	(NH4)6Mo7O24 x 4 H2O	
99	mg	FeSO4 x 7 H2O	
50	ml	**"Metals 44"**	
950	ml	Distilled water	

1. Dissolve the nitrilotriacetic acid.
2. Adjust pH to 7.0 with KOH (~7.3 g).
3. Dissolve other salts separately and combine.
4. Adjust pH to 6.8 with NaOH or H2SO4.


## "Metals 44"

250	mg	Na-EDTA	
1095	mg	ZnSO4 x 7 H2O	
500	mg	FeSO4 x 7 H2O	
154	mg	MnSO4 x H2O	
39.2	mg	CuSO4 x 5 H2O	
24.8	mg	Co(NO3)2 x 6 H2O	
17.7	mg	Na2B4O7 x 10 H2O	
1000	ml	Distilled water	

1. Dissolve the EDTA and add a few drops of concentrated H2SO4 to retard precipitation of the heavy metal ions.