# 1600: FRANCISELLA PERSICA MEDIUM


## Main sol. 1600

1. 2x concentrated stock:

1	pack	L-15 powder (Leibovitz medium)	
0.299	g	DL-aspartic acid	
0.5	g	L-Glutamic acid	
10	ml	L-Glutamine (200 mM)	
0.3	g	L-Proline	
0.299	g	alpha-ketoglutaric acid	
2.239	g	D-Glucose	
1	ml	**Stock Solution D (10 ml)**	
1	ml	**Vitamin Stock (10 ml)**	
346	ml	Demineralized water	

2. Adjust pH to 6.8 with NaOH.
3. When ready to use add: 
4. Aliquot 17.3ml into 50ml tubes and store at -20°C.
5. Fill up to 346ml with demineralized water and filter sterilize through 0.2µm filter

25	ml	Demineralized water (sterile )	
5	ml	Tryptose-phosphate (broth)	
2.5	ml	FBS (heat inactivated)	
200	µl	cholesterol lipid concentrate (250x, ThermoFisher Scientific)	

6. Filter sterilize each tube through 0.2µm filter.


## Stock Solution A (10 ml)

0.2	g	ZnSO4 x 7 H2O	
0.16	g	MnSO4 x 1 H2O	
0.02	g	CuSO4 x 5 H2O	
0.02	g	CoCl2 x 6 H2O	


## Stock Solution B (10 ml)

0.02	g	Na2 MoO4 x 2 H2O	


## Stock Solution C (10 ml)

0.02	g	Na2SeO3	


## Stock Solution D (10 ml)

1	g	Glutathione (reduced)	
1	g	Ascorbic acid	
0.05	g	FeSO4 x 7 H2O	
0.1	ml	**Stock Solution A (10 ml)**	
0.1	ml	**Stock Solution B (10 ml)**	
0.1	ml	**Stock Solution C (10 ml)**	


## Vitamin Stock (10 ml)

0.1	g	Para-aminobenzoic acid	
0.05	g	Vitamin B12	
0.01	g	D-Biotin