# 462a: PLATES WITH FLUORANTHENE


## Main sol. 462a

1000	ml	**Main sol. 462**	
x		Fluoranthene (dissolved in acetone)	

1. Pour plates of medium 462. Dissolve 2 mg/ml fluoranthene in acetone and spread 1 ml of this solution on the surface of every pre-dried plate. Allow the acetone to evaporate under sterile conditions. Inoculate as usual.


## Main sol. 462

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**	
2.5	ml	**Vitamin solution** (Schlegel)	
1000	ml	Distilled water	

1. Adjust pH to 6.9.
2. Rehydrate and cultivate lyophilized cells in the complex medium recommended for the specific strain (e.g. medium 1, 220 or 535). After this reactivation, cultivate in mineral medium 462 with the appropriate carbon source.

1000	ml	**Main sol. 1**	
1000	ml	**Main sol. 220**	
1000	ml	**Main sol. 535**	


## 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	


## Vitamin solution

1	mg	p-Aminobenzoate	
0.2	mg	Biotin	
2	mg	Nicotinic acid	
1	mg	Thiamine-HCl x 2 H2O	
0.5	mg	Calcium pantothenate	
5	mg	Pyridoxamine	
2	mg	Vitamin B12	
100	ml	Distilled water	


## Main sol. 1

5	g	Peptone	
3	g	Meat extract	
15	g	Agar, for solid medium	
1000	ml	Distilled water	

1. Adjust pH to 7.0.
2. For Bacillus strains the addition of 10.0 mg MnSO4 x H2O is recommended for sporulation.


## Main sol. 220

15	g	Casein peptone	
5	g	Soy peptone	
5	g	NaCl	
15	g	Agar	
1000	ml	Distilled water	

1. Adjust pH to 7.3. Medium is identical with Tryptone Soya Agar (Oxoid CM 131).


## Main sol. 535

30	g	Trypticase soy broth (BBL 11768, Oxoid CM129 or Merck 5459)	
20	g	Agar	
1000	ml	Distilled water	

1. pH 7.3
2. Autoclave at 121°C for 15 min.


## 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