# 1266: MEDIUM VL55


## Main sol. 1266

1000	ml	**Liquid media**	
1000	ml	**Solid media**	


## Liquid media

1.95	g	MES (2-Morpholinoethanesulfonic acid)	
10	ml	MgSO4 x 7 H2O (20 mM)	
10	ml	CaCl2 x 2 H2O (30 mM)	
10	ml	(NH4)2HPO4 (20 mM)	
1	ml	**Selenite-tungstate solution**	
1	ml	**Trace element solution SL-10**	
960	ml	Distilled water	

1. Adjust to pH 5.5 with NaOH/KOH solution (see below).
2. After autoclaving add: 

3	ml	**Vitamin solution**	
10	ml	Glucose (filter-sterilized, 0.2M)	


## Solid media

3.9	g	MES (2-Morpholinoethanesulfonic acid)	
20	ml	MgSO4 x 7 H2O (20 mM)	
20	ml	CaCl2 x 2 H2O (30 mM)	
20	ml	(NH4)2HPO4 (20 mM)	
2	ml	**Selenite-tungstate solution**	
2	ml	**Trace element solution SL-10**	
920	ml	Distilled water	

1. Adjust to pH 5.5 with NaOH/KOH solution (see below).
2. After autoclaving, keep at about 60°C, then add to each 250 ml lot: 

1.5	ml	**Vitamin solution**	
10	ml	Glucose (filter-sterilized, 0.2M)	
250	ml	Agar (autoclaved, kept about 60°C, 3%, washed)	

3. Autoclave in 250ml lots, with a magnetic stirring bar in each bottle.


## 20 mM MgSO4 x 7 H2O

100	ml	H2O	
0.49	g	MgSO4 x 7 H2O	


## 30 mM CaCl2 x 2 H2O

100	ml	H2O	
0.44	g	CaCl2 x 2 H2O	


## 20mM (NH4)2HPO4

100	ml	H2O	
0.26	g	(NH4)2HPO4	


## NaOH/KOH

100	ml	H2O	
0.8	g	NaOH	
0.56	g	KOH	


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	


## Trace element solution SL-10

10	ml	HCl (25%)	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
990	ml	Distilled water	

1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.


## Vitamin solution

17	mg	Vitamin B12	
13	mg	4-aminobenzoate	
3	mg	Biotin	
33	mg	Nicotinic acid	
17	mg	Hemicalcium D-(+)-pantothenate	
50	mg	Pyridoxamine hydrochloride	
33	mg	Thiamine-HCl x 2 H2O	
10	mg	DL-6,8-thioctic acid	
10	mg	Riboflavin	
4	mg	Folic acid	
1000	ml	Distilled water	

1. Filter sterilize the solution.