# 289: RUMINICLOSTRIDIUM PAPYROSOLVENS MEDIUM


## Main sol. 289

200	ml	Sea water (filtered)	
150	ml	**Mineral salt solution**	
1	g	NH4Cl	
0.6	g	Yeast extract	
1.65	g	K2HPO4	
0.5	ml	Sodium resazurin (0.1% w/v)	
0.5	g	L-Cysteine HCl x H2O	
5	g	Cellobiose	
5	g	Cellulose, Lens tissue or MN 301 (optional)	
650	ml	Distilled water	

1. Dissolve ingredients (except cysteine and cellobiose) and sparge medium with 100% N2 gas for 30 - 45 m in to make it anoxic. Add cysteine, adjust pH to 7.2 and dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add cellobiose from an anoxic stock solution sterilized by filtration.
2. Note: Some strains can be adapted to cellulose as substrate using 5.0 g/l cellulose (Lens tissue, OXOID; or cellulose powder MN 301, MACHEREY-NAGEL). If necessary adjust pH of completed medium to 7.2.


## Mineral salt solution

6	g	(NH4)2SO4	
6	g	NaCl	
1.2	g	MgSO4 x 7 H2O	
0.8	g	CaCl2 x 2 H2O	
1000	ml	Distilled water