# 465g: MINERAL MEDIUM WITH CYANURIC ACID AS NITROGEN SOURCE


## Main sol. 465g

3.5	g	Na2HPO4 x 2 H2O	
1	g	KH2PO4	
0.1	g	MgCl2 x 6 H2O	
1	ml	**Trace element solution SL-4**	
1000	ml	Distilled water	

1. Prepare and autoclave medium. Add 5.5 g/l glycerol from a sterile solution. Add 645 mg/l cyanuric acid from a neutralized and filter sterilized stock solution and 10 ml vitamin solution (see medium 461) to the medium.

5.5	g/l	Glycerol	
645	mg/l	Cyanuric acid	
10	ml	**Vitamin solution**	

2. Rehydrate and cultivate lyophilized cells in the complex medium recommended for the specific strain (e.g. medium 1, 220, 535 or 830). After this reactivation, cultivate on mineral medium 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-4

0.5	g	Na2-EDTA	
0.2	g	FeSO4 x 7 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 2 H2O	
0.02	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds.