# J989: IRD AMINOBACTERIUM MEDIUM


## Main sol. J989

1	g	NH4Cl	
0.3	g	KH2PO4	
0.3	g	K2HPO4	
20	g	NaCl	
0.1	g	KCl	
0.1	g	CaCl2 x 2 H2O	
1	g	L-Serine	
2	g	Yeast extract (BD-Difco)	
1	ml	**Trace element solution**	
1	ml	**Selenite-tungstate solution**	
0.5	g	L-Cysteine HCl x H2O	
0.5	mg	Resazurin	
945	ml	Distilled water	

1. Mix components thoroughly and adjust pH to 7.2. Bring to a boil, cool down under a N2-CO2 (4:1, v/v) gas stream, seal with butyl rubber stoppers and autoclave. Aseptically and anaerobically add the following stock solutions [autoclaved and stored under a N2 atmosphere or *filter-sterilize and store under a N2-CO2 (4:1, v/v) gas mixture]: 

20	ml	MgCl2 x 6 H2O (15%)	
25	ml	NaHCO3 (*, 8%)	
10	ml	Na2S x 9 H2O (5%)	

2. Readjust pH to 7.2, if necessary.


## 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