# 1478: NATRANAEROARCHAEUM MEDIUM AARC1


## Main sol. 1478

72	g	NaCl	
2.5	g	K2HPO4	
0.125	g	(NH4)2SO4	
142.5	g	Na2CO3	
22.5	g	NaHCO3	
1	ml	**Trace element solution**	
2	g	Sulfur powder	
1000	ml	Distilled water	

1. Prepare the liquid medium. Dispense the medium into Hungate tubes or serum bottles under gassing with N2. Steam medium for 3h on each of 3 successive days. Before inoculation add from sterile stock solutions:

1.2	mM	MgCl2 x 6 H2O	
1.25	mM	NaHCO3	
3	mM	NH4Cl	
50	mM	Sodium formate	
2	mM	Sodium acetate	
20	mg/l	Yeast extract	
1	ml	**Vitamine mixture**	
1	mM	Disodium sulfide (1M filter sterilized stock)	


## 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