# 1615: Basic mineral medium


## Main sol. 1615

1000	ml	**20 mM K-P-Buffer**	

1. After sterilization add from sterile stock solutions:

1	ml	MgSO4 x 7 H2O (1 M)	
10	ml	Sodium acetate (1 M)	
1	ml	CaCl2 x 2 H2O (5%)	

2. might result in colloid formation, especially when added into
hot medium, but dissolves after vigorous shaking

1	ml	Na-aspartate (2 M)	
1	ml	**Trace element solution**	
1	ml	L-Methionine (50 mM, filter sterilized, neutralised)	
0.5	ml	**Vitamine mixture**	

3. pH 7.0-7.2


## Trace element solution

5	g	EDTA	
2	g	FeSO4 x 7 H2O	
0.1	g	ZnSO4	
0.03	g	MnCl2	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 5 H2O	
0.02	g	NiCl2 x 6 H2O	
0.02	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Adjust pH to 3.0-4.0 with HCl.
2. Autoclave at 121 °C for 15 min.
3. Note: autoclave in closed bottles


## 20 mM K-P-Buffer

130	ml	K2HPO4 (0.1 M)	
70	ml	KH2PO4 (0.1 M)	
800	ml	Distilled water	

1. Sterilize at 121°C for 20 minutes in 1L Schott bottle


## Vitamine mixture

100	ml	**Acidic thiamine / Ca pantothenate solution**	
100	ml	**Basic vitamin solution**	
100	ml	**Neutral folic acid / riboflavin solution**	
100	ml	**Neutral B12 solution 0.005 g/L**	

1. Sterilize by filtration.
2. Mix the four solutions in the ratio 1:1:1:1.


## Acidic thiamine / Ca pantothenate solution

5	mg	Thiamine	
5	mg	Calcium pantothenate	
100	ml	Distilled water (acidified to pH 3.0 with HCl)	


## Basic vitamin solution

2	mg	Biotin	
5	mg	PABA (p-aminobenzoic acid)	
5	mg	Nicotinic acid	
10	mg	Pyridoxine	
100	ml	NaOH (0.1N)	


## Neutral folic acid / riboflavin solution

2	mg	Folic acid	
5	mg	Riboflavin	
100	ml	Distilled water	


## Neutral B12 solution 0.005 g/L

0.5	mg	B12	
100	ml	Distilled water