# J1309: 3.8M CHARACTERIZATION MEDIUM NO. 1


## Main sol. J1309

830	ml	Salt water (see below, 30%)	
0.025	g	Peptone (Oxoid)	
5	mg	Yeast extract	

1. Mix components and autoclave. After cooling, add the following solutions (autoclaved or *filter-sterilized): 

5	ml	NH4Cl (1M)	
2	ml	K2HPO4 (0.5 M)	
5	ml	FeSO4 x 7 H2O (*, 0.15%)	
5	ml	**Trace element solution**	
3	ml	**Vitamin solution** (*)	
50	ml	Sodium pyruvate (1.0 M)	
50	ml	Sodium perchlorate (1.0 M)	
50	ml	Sodium Thiosulfate (1.0 M)	

2. 30% Salt water: 

240	g	NaCl	
24.9	g	MgCl2 x 6 H2O	
29	g	MgSO4 x 7 H2O	
7	g	KCl	
5	ml	CaCl2 (1M)	

3. Aseptically distribute the medium into culture vessels (e.g., 10 ml medium in Balch tubes), seal with butyl rubber stoppers and replace the gas phase with N2.
4. Add components to distilled water and bring volume to 1.0 L. Adjust pH to 7.5.


## Vitamin solution

33	mg	Thiamine HCl	
10	mg	Riboflavin	
33	mg	Nicotinic acid	
17	mg	DL-Calcium pantothenate	
17	mg	Vitamin B12	
13	mg	p-Aminobenzoic acid	
1000	ml	Distilled water	


## Trace element solution

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	
1000	ml	Distilled water