# 461c: MINERAL MEDIUM WITH O-NITROPHENOL


## Main sol. 461c

800	ml	**Main sol. 461**	
200	ml	**Nitrophenol solution**	

1. Prepare and autoclave medium 461 (vitamins not necessary). Add 200 ml nitrophenol solution to 800 ml medium 461.


## Main sol. 461

1000	ml	Distilled water	
1.45	g	Na2HPO4 x 2 H2O	
0.25	g	KH2PO4	
0.01	g	CaCl2	
0.5	g	MgSO4 x 7 H2O	
0.01	g	MnSO4	
0.3	g	NH4Cl	
0.05	g	NaCl	
5	ml	**Vitamin solution**	
1	ml	**Trace element solution SL-10**	

1. Dissolve and autoclave the phosphates separately. Add the vitamin solution and trace element solution after autoclaving. After mixing, pH should be 7.5.
2. Rehydrate and cultivate the lyophilized cells in complex medium recommended for the specific strain (e.g. medium 1, 220 or 535) first. After this reactivation, cultivate on mineral medium 461 with the appropriate carbon source.


## Nitrophenol solution

0.5	g	2-Nitrophenol	
1000	ml	Phosphate buffer (50 mM, pH 7.5)	

1. Dissolve 0.5 g o-nitrophenol per liter phosphate buffer (50 mM, pH 7.5) and sterilize by filtration.