# J1301: SPIROCHAETEA NS MEDIUM


## Main sol. J1301

1. Mix components thoroughly, adjust pH to 8.0, bring to a boil for a few seconds and cool down under a stream of N2. Distribute the medium into culture vessels (e.g., 10 ml in Hungate tubes) under a N2 gas stream, seal with butyl rubber stoppers and autoclave. After cooling, aseptically and anaerobically add per liter the following solutions from sterile anaerobic stocks (autoclaved or *filter-sterilized): 

1	g	NaNO3	
0.2	g	K2HPO4	
0.25	g	NH4Cl	
0.4	g	MgSO4 x 7 H2O	
0.5	g	FeSO4 x 7 H2O	
0.11	g	CaCl2 x 2 H2O	
1	ml	**Trace mineral solution**	
1	mg	Resazurin	
1000	ml	Distilled water	
20	ml	**Trace vitamins** (*)	
10	ml	Maltose (1.0 M)	
5	ml	Na2S x 9 H2O (5%)	


## Trace mineral solution

1000	ml	**Trace minerals**	
0.03	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
0.4	mg	Na2WO4 x 2 H2O	


## 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