# 602: HALOMONAS SUBGLACIESCOLA


## Main sol. 602

80	g	NaCl	
9.5	g	MgSO4 x 7 H2O	
0.5	g	KCl	
0.2	g	CaCl2 x 2 H2O	
0.1	g	(NH4)2SO4	
0.1	g	KNO3	
1	g	Yeast extract	
960	ml	Distilled water	
20	ml	**Hutner's salts**	
1	ml	**Vitamin solution**	
20	ml	**Phosphate supplement**	

1. pH 8.0
2. Autoclave and add, on cooling 20.0 ml sterile Hutner's salts (see medium 590), 1.0 ml vitamin solution, and 20.0 ml sterile phosphate supplement.


## Vitamin solution

10	mg	Vitamin B12	
2	mg	Biotin	
10	mg	Thiamine-HCl x 2 H2O	
5	mg	Calcium pantothenate	
2	mg	Folic acid	
5	mg	Nicotinamide	
10	mg	Pyridoxine hydrochloride	
1000	ml	Distilled water	

1. Filter-sterilized.


## Phosphate supplement

50	mg	K2HPO4	
50	mg	KH2PO4	
20	ml	Distilled water	


## 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.