# 1266a: MEDIUM GY VL55


## Main sol. 1266a

500	ml	**Solution 1**	
500	ml	**Solution 2**	

1. After autoclaving, cool to 50-60°C and combine the solution by pouring solution 2 to solution 1.


## Solution 1

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

1. Adjust to pH 5.5 with NaOH/KOH solution (see below).


## Solution 2

1	g	Glucose	
0.5	g	Yeast extract	
500	ml	Distilled water	
20	g	Agar, for solid medium	


## 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.