# J1245: TRICHOCOCCUS SKBG MEDIUM


## Main sol. J1245

1. Mix components thoroughly and adjust pH to 7.2. Distribute the medium into culture vessels (e.g., 10 ml in Hungate tubes) under a stream of N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. Allow to stand overnight. Aseptically and anaerobically add the following solutions (filter-sterilized): 

1	g	NaCl	
0.2	g	MgCl2 x 6 H2O	
0.91	g	NaH2PO4 x 2 H2O	
0.1	g	CaCl2 x 2 H2O	
1	g	NH4Cl	
0.2	g	Yeast extract (BD-Difco)	
1	ml	**FeCl2 solution**	
1	ml	**Trace element solution**	
900	ml	Distilled water	
30	ml	NaHCO3 (8.0%)	
10	ml	**Trace vitamins**	
50	ml	Lactose (0.25 M)	

2. Readjust pH to 7.2-7.6, if necessary.


## 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