# 1073: AML60 MEDIUM


## Main sol. 1073

500	ml	**Solution 1**	
500	ml	**Solution 2**	

1. Dissolve ingredients (except carbonates, vitamins, arsenate and sulfide), boil medium for 1 min., then cool to room temperature under N2 gas atmosphere. Add solid carbonates and adjust pH to 9.8, then dispense under same gas atmosphere in culture vessels and autoclave.
2. Prior to inoculation, add vitamins, arsenate and sulfide from sterile, anaerobic stock solutions prepared under N2 and check pH of the completed medium. The vitamin solution should be sterilized by filtration.
3. Growth of strain DSM 17814 takes place without visible increase in turbidity, hence proliferation of cells has to be checked using phase contrast microscopy.


## Solution 1

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	**Selenite-tungstate solution**	
500	ml	Distilled water	


## Solution 2

10.6	g	Na2CO3	
4.2	g	NaHCO3	
500	ml	Distilled water	


## 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.


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water