# 1024: NITRATIRUPTOR AND NITRATIFRACTOR MEDIUM


## Main sol. 1024

1	g	NaNO3	
1	g	NaHCO3	
1	g	Na2S2O3 x 5 H2O	
3	g	Sulfur powder	
10	ml	**Wolin's vitamin solution**	
1000	ml	**Synthetic seawater**	

1. Prepare the medium under an atmosphere of H2/CO2 (80:20) without adding the vitamins and NaHCO3 in serum bottles and seal the serum tubes with butyl rubber stoppers. Steam medium for 3 hours on each of 3 successive days. To the sterile medium add, from filter sterilised stock solutions, the NaHCO3 and vitamin solution.
2. Increase the 80% H2 + 20% CO2 gas phase pressure to 300 kPa. The final pH is 7.0.


## 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	


## 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	Nitrilotriacetic acid	
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