# J823: TEPIDIBACTER MESOPHILUS MEDIUM


## Main sol. J823

5	g	NaCl	
4	g	MgCl2 x 6 H2O	
0.34	g	KCl	
0.25	g	NH4Cl	
0.11	g	CaCl2 x 2 H2O	
0.18	g	K2HPO4	
2	mg	Fe(NH4)2(SO4)2 x 6 H2O	
1	ml	**FeCl2 solution**	
1	ml	**Trace element solution**	
0.2	g	Yeast extract	
2	g	Casitone (BD-Difco)	
10	ml	**Trace vitamins**	
1	mg	Resazurin	
1000	ml	Distilled water	

1. Mix components and bring to a boil for 5-10 sec and cool down under a N2-CO2 (80:20, v/v) gas stream. Dispense the medium into culture vessels (e.g., 20 ml in 50 ml serum bottles) under the same gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, add per liter the following solutions from sterile anaerobic stocks: 

10	ml	Na2S x 9 H2O (5%)	
5	ml	NaHCO3 (8%)	

2. To prepare the stock solutions, autoclave 5% Na2S x 9H2O solution under a N2 atmosphere, filter-sterilize and store 8% NaHCO3 solution under a N2-CO2 (80:20, v/v) gas mixture.


## FeCl2 solution

10	ml	HCl (7.7 M, 25%)	
1.5	g	FeCl2 x 4 H2O	
990	ml	Distilled water	


## Trace element solution

70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 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