# J1078: ALKALIPHILUS 3B MEDIUM


## Main sol. J1078

1	g	NaCl	
1	g	NH4Cl	
0.1	g	KH2PO4	
0.1	g	K2HPO4	
0.1	g	KCl	
0.1	g	MgCl2 x 6 H2O	
3	g	Tris base	
1.5	g	Na2S2O3 x 5 H2O	
1	ml	**Trace element solution**	
1	mg	Resazurin	
910	ml	Distilled water	

1. Mix components thoroughly, adjust pH to 9.0 with NaOH and autoclave under a N2 atmosphere. After cooling, replace the gas phase with a N2-CO2 (4:1, v/v) gas mixture. Add the following solutions from sterile anaerobic stocks (autoclaved and stored under a N2 atmosphere): 

50	ml	Yeast extract (w/v, 10%)	
20	ml	Crotonic acid (pH 7.0, 0.5 M)	
20	ml	Na2CO3 (8%)	
6	ml	Na2S x 9 H2O (5%)	

2. Check final pH be around 8.5.


## 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