# 1230: THIOALKALIMICROBIUM MICROAEOPHILUM MEDIUM


## Main sol. 1230

6	g	NaCl	
2	g	K2HPO4	
40	g	NaHCO3	
0.5	g	KNO3	
1000	ml	Distilled water	

1. Adjust the pH to 9.0 - 9.5 with 2M KOH. Dispense into closed vessels (i.e. a serum tube or bottle) with a liquid to air ratio of about 1:10. Flush the gas phase with nitrogen and add 2% oxygen. Sterilize at 120°C 20 min.
2. After sterilization add from sterile stock solutions:

1	ml	**Trace element solution**	
1	ml	MgCl2 x 6 H2O (200.0 g/l)	

3. (a white colloid will form which will rapidly dissolve after mixing)

5	ml	Sodium thiosulfate pentahydrate (4 M - 992 g/l)	

4. Incubate statically until growth occurs, then shake at 100 rpm.


## Trace element solution

5	mg	EDTA	
2	mg	FeSO4 x 7 H2O	
100	mg	ZnSO4 x 7 H2O	
30	mg	MnCl2 x 4 H2O	
200	mg	CoCl2 x 6 H2O	
20	mg	NiCl2 x 6 H2O	
30	mg	Na2MoO4 x 2 H2O	
10	mg	CuCl2 x 2 H2O	
300	mg	H3BO3	
1000	ml	Distilled water	

1. Final pH should be 3, add HCl if needed. Sterilization - 120°C 20 min.