# J1029: RUMEN BACTERIA MEDIUM


## Main sol. J1029

38	ml	**Mineral solution**	
0.3	g	K2HPO4	
2	g	Trypticase peptone (BD-BBL)	
0.5	g	Yeast extract	
3.1	ml	**VFA solution**	
1	mg	Hemin	
0.5	g	Glucose	
0.5	g	Maltose	
0.5	g	Cellobiose	
0.5	g	Starch,  soluble	
0.5	g	Glycerol	
4	g	Na2CO3	
1	mg	Resazurin	
0.25	g	L-Cysteine HCl x H2O	
0.25	g	Na2S x 9 H2O	
960	ml	Distilled water	

1. Adjust pH to 6.7 - 6.8; gas atmosphere: 100% CO2.
2. Dissolve ingredients except carbohydrates, Na2CO3, cysteine and sodium sulfide in distilled water, boil for 5 min, then cool to room temperature while gassing with 100% CO2. Add Na2CO3 and equilibrate the medium with the gas to pH 6.8. Distribute under CO2 in rubber-stoppered tubes (i.e. Hungate tubes) and autoclave. Thereafter add carbohydrate mixture, cysteine and sodium sulfide from concentrated sterile stock solutions (prepared anaerobically under N2 gas and autoclaved).


## Mineral solution

6	g	KH2PO4	
12	g	NaCl	
6	g	(NH4)2SO4	
1.6	g	CaCl2 x 2 H2O	
2.5	g	MgSO4 x 7 H2O	
1000	ml	Distilled water	


## VFA solution

17	ml	Acetic acid	
6	ml	Propionic acid	
4	ml	n-Butyric acid	
1	ml	iso-Butyric acid	
1	ml	n-Valeric acid	
1	ml	iso-Valeric acid	
1	ml	DL-alpha-Methyl butyric acid