# 997: HNW MEDIUM


## Main sol. 997

1	g	NaNO3	
1	g	NaHCO3	
0.1	mg	Na2WO4 x 2 H2O	
0.5	g	Na2S	
10	ml	**Wolin's vitamin solution**	
1000	ml	**Synthetic seawater**	
100	g/l	NaNO3	
100	g/l	NaHCO3	
100	g/l	Na2S	

1. To prepare the medium, all compounds of DMJ seawater were dissolved in 1 liter of distilled deionized water, and the pH was adjusted to around 7.0 with NaOH at room temperature prior to autoclaving. After autoclaving, filter-sterilized NaNO3 solution (100 g/l), NaHCO3 solution (100 g/l), Na2S solution (100 g/l); pH 7.5) and trace vitamin solution were added. Then the tubes were tightly sealed with butyl rubber stoppers under a gas phase of 80% H2 + 20% CO2 (300 kPa).


## Synthetic seawater

30	g	NaCl	
0.14	g	K2HPO4	
0.14	g	CaCl2 x 2 H2O	
0.25	g	NH4Cl	
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	**Trace mineral solution**	


## Trace mineral solution

1.5	g	C6H9NO6	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x 2 H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.18	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.18	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.02	g	AlK(SO4)2 x 12 H2O	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
0.025	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
1000	ml	Distilled water	


## Wolin's vitamin solution

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 D-(+)-pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water