# J1093: MJ BASAL MEDIUM WITH THIOSULFATE/2


## Main sol. J1093

1000	ml	**MJ(-N) synthetic seawater**	
0.25	g	NH4Cl	
0.25	g	KNO3	
1000	ml	Distilled water	

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

10	ml	NaHCO3 (8%)	
15	ml	Na2S2O3 x 5 H2O (10%)	
1	ml	**Trace vitamins**	

2. Aseptically distribute the medium into culture vessels (e.g., 20 ml in 120 ml serum bottles) and seal with butyl rubber stoppers. Replace the gas phase with a N2-CO2 (80:20, v/v) gas mixture, and pressurize to 200 kPa. Adjust pH to 6.0, if necessary.


## MJ(-N) synthetic seawater

30	g	NaCl	
0.14	g	K2HPO4	
0.14	g	CaCl2 x 2 H2O	
3.4	g	MgSO4 x 7 H2O	
4.18	g	MgCl2 x 6 H2O	
0.33	g	KCl	
0.5	mg	NiCl2 x 6 H2O	
0.5	mg	Na2SeO3 x 5 H2O	
0.01	g	Fe(NH4)2(SO4)2 x 6 H2O	
10	ml	**Modified Wolin's mineral solution**	
1000	ml	Distilled water	


## 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	


## Trace minerals

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x n H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.1	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.01	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 solution. Then proceed to add minerals. Adjust final pH to 7.0.