# J1149: METHANOMASSILIICOCCUS ALVUS MEDIUM


## Main sol. J1149

0.5	g	KH2PO4	
0.4	g	MgSO4 x 7 H2O	
5	g	NaCl	
1	g	NH4Cl	
0.05	g	CaCl2 x 2 H2O	
2.7	g	Sodium acetate x 3 H2O	
1	ml	**Trace element solution**	
0.5	g	L-Cysteine HCl x H2O	
1	ml	**Selenite-tungstate solution**	
1	mg	Resazurin	
980	ml	Distilled water	

1. Mix components and adjust pH to 7.0. Bring to a boil and cool down under a H2-CO2 (4:1, v/v) gas stream. Distribute the medium into culture vessels under the same gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, aseptically and anaerobically add per liter the following solutions (autoclaved or *filter-sterilized): 

20	ml	NaHCO3 (*, 8%)	
8	ml	Na2S x 9 H2O (5%)	


## Trace element solution

12.5	ml	HCl (25%)	
2.1	g	FeSO4 x 7 H2O	
30	mg	H3BO3	
100	mg	MnCl2 x 4 H2O	
190	mg	CoCl2 x 6 H2O	
24	mg	NiCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
144	mg	ZnSO4 x 7 H2O	
36	mg	Na2MoO4 x 2 H2O	
987	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