# 1840: Halorutilales haloarchaeon medium


## Main sol. 1840

240	g	NaCl	
2.5	g	K2HPO4	
5	g	KCl	
0.2	g	NH4Cl	
1	ml	**Trace element solution**	
999	ml	Distilled water	

1. Adjust pH to 6.8 with 0.5 M KH2PO4.
2. Dispense into serum bottles (30 ml medium in 100 ml serum bottles, OR 15 ml in 50 serum bottles).
3. add 20% CO2 to gas phase (in serum bottles)
4. Autoclave at 121 °C for 20 min.
5. Before inoculation add from sterile stock solutions per liter:

0.5	ml/l	**Vitamine mixture**	
0.08	ml/l	MgSO4 x 7 H2O (1M in water)	
0.5	ml/l	**alkaline trace metal solution**	
10	ml/l	Na-thiosulfate (2M in water)	
10.5	ml/L	NaHCO3 (8% in water)	


## Trace element solution

5	g	EDTA	
2	g	FeSO4 x 7 H2O	
0.1	g	ZnSO4	
0.03	g	MnCl2	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 5 H2O	
0.02	g	NiCl2 x 6 H2O	
0.02	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Adjust pH to 3.0-4.0 with HCl.
2. Autoclave at 121 °C for 15 min.
3. Note: autoclave in closed bottles


## Vitamine mixture

100	ml	**Acidic thiamine / Ca pantothenate solution**	
100	ml	**Basic vitamin solution**	
100	ml	**Neutral folic acid / riboflavin solution**	
100	ml	**Neutral B12 solution 0.005 g/L**	

1. Sterilize by filtration.
2. Mix the four solutions in the ratio 1:1:1:1.


## Acidic thiamine / Ca pantothenate solution

5	mg	Thiamine	
5	mg	Calcium pantothenate	
100	ml	Distilled water (acidified to pH 3.0 with HCl)	


## Basic vitamin solution

2	mg	Biotin	
5	mg	PABA (p-aminobenzoic acid)	
5	mg	Nicotinic acid	
10	mg	Pyridoxine	
100	ml	NaOH (0.1N)	


## Neutral folic acid / riboflavin solution

2	mg	Folic acid	
5	mg	Riboflavin	
100	ml	Distilled water	


## Neutral B12 solution 0.005 g/L

0.5	mg	B12	
100	ml	Distilled water	


## alkaline trace metal solution

0.04	g/l	NaOH	
0.026	g/l	Na2SeO3 x 5 H2O	
0.03	g/l	Na2WO4 x 2 H2O	
1000	ml	Distilled water