# J874: BELH1 MEDIUM


## Main sol. J874

0.1	g	KH2PO4	
0.1	g	K2HPO4	
1	g	NH4Cl	
70	g	NaCl	
0.1	g	KCl	
0.1	g	CaCl2 x 2 H2O	
10	ml	**Trace element solution**	
1	g	Yeast extract	
5	g	Na2S2O3 x 5 H2O	
1	mg	Resazurin	
935	ml	Distilled water	

1. Mix components thoroughly, adjust pH to 7.5 with KOH and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions (autoclaved and stored under N2, or *filter-sterilized and stored under N2-CO2, 4:1, v/v): 

20	ml	Glucose (1 M)	
25	ml	NaHCO3 (*, 8%)	
20	ml	MgCl2 x 6 H2O (15%)	

2. Aseptically distribute the medium into culture vessels under the same gas mixture and seal with butyl rubber stoppers. Finally, add per liter the following solutions (autoclaved and stored under N2): 

10	ml	L-Cysteine HCl x H2O (5%)	
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