# 591: HALOBACTEROIDES/HALOINCOLA MEDIUM


## Main sol. 591

0.33	g	NH4Cl	
0.33	g	CaCl2	
0.33	g	MgCl2	
0.33	g	KCl	
0.33	g	KH2PO4	
125	g	NaCl, adjust if required in range 100-150	
1.5	g	NaHCO3	
0.5	g	Na2S x 9 H2O	
5	g	Glucose	
5	g	Peptone (Bacto Proteose Peptone No.3)	
1	ml	**Trace element solution SL-10** (SL 10)	
10	ml	**Wolin's vitamin solution**	
2	mg	Resazurin	
990	ml	Distilled water	

1. Prepare the medium without NaHCO3, Na2S, glucose and vitamins, boil and cool under a nitrogen:carbon dioxide (80:20 v/v) atmosphere. Glucose (filter-sterilized and stored under nitrogen), vitamins (filter sterilized and stored under nitrogen), and sodium sulphide (autoclaved under nitrogen) are added from sterile stock solutions. The pH should be pH 7.5.


## 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

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium D-(+)-pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water