# J1162: NAS-02 MEDIUM


## Main sol. J1162

0.2	g	(NH4)2SO4	
3	g	KH2PO4	
1	ml	**Modified Allen's trace metal solution**	
2.5	g	Na2S2O3 x 5 H2O	
1000	ml	Distilled water	

1. Mix components thoroughly and adjust pH to 5.2. Autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions anaerobically and aseptically (autoclaved or *filter-sterilized and store under a N2 atmosphere): 

10	ml	MgSO4 x 7 H2O (w/v, 5%)	
5	ml	CaCl2 x 2 H2O (w/v, 5%)	
10	ml	**Trace vitamins** (*)	
5	ml	Yeast extract (w/v, 10%)	
10	ml	L-Cysteine HCl (25 mM)	
10	ml	FeCl2 (130 mM)	


## Modified Allen's trace metal solution

1.8	g	MnCl2 x 4 H2O	
4.5	g	Na2B4O7 x 10 H2O	
0.22	g	ZnSO4 x 7 H2O	
0.05	g	CuCl2 x 2 H2O	
0.03	g	Na2MoO4 x 2 H2O	
0.03	g	VOSO4 x n H2O	
0.01	g	CoSO4 x 7 H2O	
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