# J1175: CAENICOLA MOBILE MEDIUM


## Main sol. J1175

0.5	g	KH2PO4	
0.5	g	K2HPO4	
0.5	g	NH4Cl	
0.4	g	NaCl	
0.3	g	MgCl2 x 6 H2O	
0.05	g	CaCl2 x 2 H2O	
0.4	g	KCl	
2	g	Yeast extract	
2	g	Tryptone (BD-Difco)	
2	g	Casamino acids (BD-Difco)	
2.4	g	L-Cysteine	
4.2	g	L-Arginine	
4.2	g	L-Histidine	
1	ml	**Trace metal solution**	
1	mg	Resazurin	
975	ml	Distilled water	

1. Mix components thoroughly, adjust pH to 7.5 with KOH and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions (filter-sterilized): 

25	ml	NaHCO3 (8%)	
1	ml	**Trace vitamins**	

2. Aseptically and anaerobically distribute the medium into culture vessels (e.g., 20 ml medium in 60 ml serum bottles) under a N2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Then, add per liter the following solution (autoclaved and stored under N2): 

8	ml	Na2S x 9 H2O (5%)	

3. Readjust pH to 7.5, if necessary.


## Trace metal solution

10	ml	HCl (32%)	
2	g	FeCl2 x 4 H2O	
0.25	g	CoCl2 x 6 H2O	
0.1	g	MnCl2 x 4 H2O	
0.07	g	ZnCl2	
6	mg	H3BO3	
0.04	g	Na2MoO4 x 2 H2O	
0.07	g	NiCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
0.025	g	AlCl3	
6	mg	Na2WO4 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