# 999: HALORHODOSPIRA MEDIUM


## Main sol. 999

1	g	KH2PO4	
0.05	g	CaCl2 x 2 H2O	
0.5	g	NH4Cl	
100	g	NaCl	
2	g	MgCl2 x 6 H2O	
1	g	MgSO4 x 7 H2O	
1000	ml	Distilled water	

1. After autoclaving 20 min at 121°C, the following sterile solutions are added:  

10	ml	NaHCO3 (20%)	
7.5	ml	Na2S x 9 H2O (10%)	
1	ml	**Vitamins V7 solution (Pfennig et Trüper, 1992)**	
1	ml	**Trace element solution SL12**	

2. The pH is adjusted to pH 7.2-7.4 For better growth of pure cultures, the growth medium should be supplemented with acetate (2 mM final concentration), succinate (1 mM final concentration) and yeast extract (0.5 g/l). Pure cultures should be incubated at 30°C and light intensity of 500 lux from a tungsten lamp (16 h light, 8 h dark). Pure cultures in liquid medium can be stored in 60 ml screw cap bottles at 4°C in the dark.


## Trace element solution SL12

1000	ml	Distilled water	
300	mg	H3BO3	
1.1	g	FeSO4 x 7 H2O	
190	mg	CoCl2 x 6 H2O	
50	mg	MnCl2 x 2 H2O	
42	mg	ZnCl2	
24	mg	NiCl2 x 6 H2O	
18	mg	Na2MoO4 x 2 H2O	
2	mg	CuCl2 x 2 H2O	

1. Sterilized by filtration.


## Vitamins V7 solution (Pfennig et Trüper, 1992)

1000	ml	Distilled water	
2	mg	Biotine	
10	mg	p-Aminobenzoate	
10	mg	Thiamine	
5	mg	Pantothenate	
50	mg	Pyridoxamine	
20	mg	Vitamine B12	
20	mg	Nicotinate	

1. Sterilized by filtration.