# J1131: SALINISPIRA L21 MEDIUM


## Main sol. J1131

60	g	NaCl	
6	g	MgCl2 x 6 H2O	
1.5	g	KCl	
1	g	Na2SO4	
1	g	NH4Cl	
0.4	g	CaCl2 x 2 H2O	
0.4	g	K2HPO4	
10	ml	**Trace mineral solution**	
1	g	Yeast extract	
1	g	Trypticase peptone (BD-BBL)	
0.5	mg	Resazurin	
950	ml	Distilled water	

1. Mix components thoroughly, bring to a boil and cool down under a N2-CO2 (4:1, v/v) gas stream and autoclave under the same gas mixture. After cooling, add per liter the following solutions (autoclaved or *filter-sterilized): 

10	ml	Glucose (10%)	
10	ml	**Trace vitamins** (*)	
30	ml	NaHCO3 (*, 8%)	

2. Aseptically and anaerobically, dispense the medium into culture vessels (e.g., 10 ml in Hungate tubes) under a N2-CO2 (4:1, v/v) gas stream and seal with butyl rubber stoppers. Prior to inoculation, add per liter the following solutions (autoclaved and stored under a N2 atmosphere): 

10	ml	L-Cysteine HCl x H2O (5%)	
10	ml	Na2S x 9 H2O (5%)	

3. Readjust pH to 7.3-7.5, if necessary.


## 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