# 461: MINERAL MEDIUM (NAGEL AND ANDREESEN)


## Main sol. 461

1000	ml	Distilled water	
1.45	g	Na2HPO4 x 2 H2O	
0.25	g	KH2PO4	
0.01	g	CaCl2	
0.5	g	MgSO4 x 7 H2O	
0.01	g	MnSO4	
0.3	g	NH4Cl	
0.05	g	NaCl	
5	ml	**Vitamin solution**	
1	ml	**Trace element solution SL-10**	

1. Dissolve and autoclave the phosphates separately. Add the vitamin solution and trace element solution after autoclaving. After mixing, pH should be 7.5.
2. Rehydrate and cultivate the lyophilized cells in complex medium recommended for the specific strain (e.g. medium 1, 220 or 535) first. After this reactivation, cultivate on mineral medium 461 with the appropriate carbon source.


## Vitamin solution

50	mg	Vitamin B12	
50	mg	Pantothenic acid	
50	mg	Riboflavin	
10	mg	Pyridoxamine hydrochloride	
20	mg	Biotin	
20	mg	Folic acid	
25	mg	Nicotinic acid	
25	mg	Nicotine amide	
50	mg	alpha-lipoic acid	
50	mg	p-Aminobenzoic acid	
50	mg	Thiamine-HCl x 2 H2O	
1000	ml	Distilled water	

1. Stir for some hours, filter sterilize the solution.


## 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.