# 457b: MEDIUM WITH FLUORANTHENE


## Main sol. 457b

1000	ml	**Main sol. 457**	
200	mg/l	Tween 80	

1. For DSM 7526, prepare medium 457 with 200 mg/l Tween 80. Prepare a solution of 2 g/l
fluoranthene in acetone and filter sterilize using a cellulose filter membrane. Add 1 ml of
this stock solution to the sterilized culture flask. Let acetone evaporate. Add 20 ml
medium to the flask. Alternatively, the direct addition of about 1-2 mg fluoranthene to
the medium is possible.


## Main sol. 457

2.44	g	Na2HPO4	
1.52	g	KH2PO4	
0.5	g	(NH4)2SO4	
0.2	g	MgSO4 x 7 H2O	
0.05	g	CaCl2 x 2 H2O	
10	ml	**Trace element solution SL-4**	
1000	ml	Distilled water	

1. Adjust pH to 6.9.
2. Prepare a separate solution of the phosphates and autoclave separately. Combine the two solutions after cooling.
3. Rehydrate and cultivate lyophilized cells in complex medium (e.g. medium 1, 220 or 535). After this reactivation, cultivate in mineral medium 457 with the appropriate carbon source.


## Trace element solution SL-4

0.5	g	EDTA	
0.2	g	FeSO4 x 7 H2O	
100	ml	**Trace element solution SL-6**	
900	ml	Distilled water	


## Trace element solution SL-6

0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 2 H2O	
0.02	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water