# 1457: ALCALILIMNICOLA EHRLICHII MHLE-1 MEDIUM


## Main sol. 1457

496	ml	**Solution A**	
500	ml	**Solution B**	
10	ml	**Solution C**	
20	g	Agar, for solid medium	

1. Autoclave solution A without the vitamin solution. Autoclave solution B in half full, closed bottles. Autoclave solution C. Combine the sterile solutions A, B, and C after cooling. Aseptically add 1.0 ml of the vitamin solution (filter sterilized) to 1 litre of the sterile medium.
2. Adjust to pH 9.8. The medium may be solidified by adding 20 g/l agar, but care should be taken that the temperature of the solutions to be mixed are about 50°C, otherwise the agar will darken and degrade.


## Solution A

0.1	g	(NH4)2SO4	
25	mg	MgSO4	
60	g	NaCl	
0.15	g	K2HPO4	
0.08	g	KH2PO4	
5	ml	**Trace element solution SL-10**	
0.3	ml	Tungstate (0.1 mM)	
1	ml	**Vitamin solution**	
490	ml	Distilled water	


## Solution B

10.6	g	Na2CO3	
4.2	g	NaHCO3	
500	ml	Distilled water	


## Solution C

10	ml	Na-acetate (1M)	


## Vitamin solution

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine-HCl x 2 H2O	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	D-Calcium pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water	


## Trace element solution SL-10

10	ml	HCl (25%; 7.7 M)	
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. Make up to a final volume of 1000.0 ml.