# J519: SYNTROPHOMONAS MEDIUM (for Syntrophospora cellicola 19J-3)


## Main sol. J519


## Solution A

50	ml	**Mineral solution**	
1	ml	**FeCl2 solution**	
1	ml	**Trace element solution**	
50	ml	Rumen fluid (see Medium No. 266),  clarified	
1	g	Trypticase peptone (BD-BBL)	
5	ml	**Vitamin solution**	
1.7	g	Butyric acid	
2.8	g	Na2SO4	
1	mg	Resazurin	
795	ml	Distilled water	

1. Mix components, adjust pH to 7.2, bring to a boil for a few seconds and cool down under a stream of a N2-CO2 (80:20) gas mixture. Distribute Solution A into culture vessels (e.g., 9 ml per tube) under the same gas mixture, seal with butyl rubber stoppers and autoclave.
2. To complete the medium, add 0.1 volume of 3.5% NaHCO3 solution [filter-sterilized and stored under a N2-CO2 (80:20) gas mixture] and reduce the medium with 0.3 g/L (final) L-cysteine x HCl x H2O and 0.3 g/L (final) Na2S x 9H2O (autoclave and store 3% stock solutions under a N2 atmosphere).


## Mineral solution

10	g	KH2PO4	
6.6	g	MgCl2 x 6 H2O	
8	g	NaCl	
8	g	NH4Cl	
1	g	CaCl2 x 2 H2O	
1000	ml	Distilled water	


## Vitamin solution

0.25	mg	Biotin	
2.5	mg	Nicotinic acid	
1.25	mg	Thiamine HCl	
1.25	mg	p-Aminobenzoic acid	
0.62	mg	DL-Calcium pantothenate	
6.2	mg	Pyridoxine hydrochloride	
1000	ml	Distilled water	


## 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