# J565: DESULFOVIBRIO HEB233 MEDIUM


## Main sol. J565


##  Solution A

0.3	g	KH2PO4	
0.3	g	K2HPO4	
1	g	NH4Cl	
3	g	Na2SO4	
23	g	NaCl	
0.1	g	KCl	
0.1	g	CaCl2	
0.1	g	Yeast extract (BD-Difco)	
1	ml	**Trace element solution**	
0.5	g	L-Cysteine HCl x H2O	
1	mg	Resazurin	
900	ml	Distilled water	

1. To complete the medium, add the following stock solutions [Filter-sterilize NaHCO3 solution and store under a N2-CO2 (4:1, v/v) gas mixture. The other solutions are autoclaved and stored anaerobically under a N2 atmosphere] to the medium (per 4.5 ml): 

0.1	ml	Sodium lactate (1 M)	
0.1	ml	MgCl2 x 6 H2O (15%)	
0.2	ml	NaHCO3 (5%)	
0.1	ml	Na2S x 9 H2O (2%)	

2. Mix ingredients and adjust pH to 7.2. Distribute 0.9 volume of the medium into tubes (e.g., 4.5 ml of the medium in the Hungate tubes) under a N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave at 110C for 45 min.


## 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