# J419: INORGANIC MINERAL MEDIUM WITH THIOSULFATE


## Main sol. J419

3	g	NaCl	
0.014	g	K2HPO4	
0.08	g	CaCl2 x 2 H2O	
0.34	g	MgSO4 x 7 H2O	
0.418	g	MgCl2 x 6 H2O	
0.33	g	KCl	
0.5	g	NH4Cl	
1	g	Na2S2O3 x 5 H2O	
0.8	g	Na2SiO3 x 9 H2O	
2	mg	Fe2(SO4)3 x n H2O	
0.1	mg	FeSO4 x 7 H2O	
0.05	mg	NiCl2 x 6 H2O	
0.05	mg	Na2SeO3 x 5 H2O	
1	ml	**Trace mineral solution**	
1	ml	**Trace vitamins**	
0.5	g	NaHCO3	
1000	ml	Distilled water	

1. Mix ingredients, except Trace vitamins and NaHCO3, and adjust pH to 7.0. Distribute the medium into culture vessels (e.g., 20 ml of medium in 120 ml serum bottles) under a stream of N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. Filter-sterilize Trace vitamins and 5% NaHCO3 solution and add to the medium. Finally add O2 gas to make 5% (v/v) in the gas phase.


## Trace mineral solution

1.5	g	Nitrilotriacetic acid	
0.5	g	MnSO4 x n H2O	
0.5	g	CoSO4 x 7 H2O	
0.18	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.02	g	AlK(SO4)2	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. Adjust final pH to 7.0.


## Trace vitamins

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water