# J1051: ENDOMICROBIUM PROAVITUM RSA MEDIUM


## Main sol. J1051

1	g	NaCl	

1. Adjust pH to 7.1 - 7.2. Distribute the medium into culture vessels under the same gas mixture and seal with butyl rubber stoppers. Finally, add per liter the following solution (filter-sterilized): 

0.5	g	KCl	
0.4	g	MgCl2 x 6 H2O	
0.1	g	CaCl2 x 2 H2O	
0.3	g	NH4Cl	
0.2	g	KH2PO4	
0.4	mg	Resazurin	
945	ml	Distilled water	

2. Mix components thoroughly and autoclave. Cool medium under a N2-CO2 (4:1, v/v) gas stream, and add the following solutions (autoclaved or *filter-sterilized) anaerobically and aseptically: 

30	ml	NaHCO3 (*, 8%)	
5	ml	Glucose (*, 1 M)	
10	ml	L-Cysteine HCl x H2O (0.1 M)	
2	ml	Yeast extract (*, 10%)	
2	ml	Casamino acids (*, 10%)	
1	ml	**Trace element solution**	
1	ml	**FeCl2 solution**	
1	ml	**Selenite-tungstate solution**	
1	ml	Trace vitamins solution (see Medium No. 284) (*)	
1	ml	**Fatty acid mixture**	
1	ml	**Aromatic fatty acids solution**	
4	ml	Dithiothreitol (250 mM)	


## Aromatic fatty acids solution

68	mg	Phenyl acetic acid	
75	mg	Phenyl propionic acid	
76	mg	4-Hydroxyphenyl acetic acid	
88	mg	3-Indolyl acetic acid	
100	ml	NaOH (20 mM)	

1. Autoclave and store under a N2 atmosphere.


## 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	


## FeCl2 solution

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


## Selenite-tungstate solution

0.4	g	NaOH	
6	mg	Na2SeO3 x 5 H2O	
8	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	


## Fatty acid mixture

0.5	g	Valeric acid	
0.5	g	Isovaleric acid	
0.5	g	alpha-Methylbutyric acid	
0.5	g	Isobutyric acid	
20	ml	Distilled water	

1. Adjust pH to 7.5 with concentrated NaOH solution.