medium no. 15: AA medium, 1 mg/L 2,4-dichlorophenoxyacetic acid, 0.2 mg/L kinetin, 1.2% (w/v) agar, pH 5.8
Plant cell lines
Chemicals and stock solutions
(All stock solutions are stored at 4°C)
Sucrose
MS_micro_1
Chemical
Concentration (mg/mL)
H3BO3
0.62
MnSO4·5H2O
2.41
ZnSO4·7H2O
0.86
KI
0.083
Na2MoO4·2H2O
0.025
CuSO4·5H2O
0.0025
CoCl2·6H2O
0.0025
MS_micro_2
Chemical
Concentration (mg/mL)
FeSO4·7H2O
2.78
Na2-EDTA
3.73
Heat at 80°C for 3–4 hours for chelating Fe
AA_1
Chemical
Concentration (mg/mL)
KCl
29.4
CaCl2·2H2O
4.4
MgSO4·7H2O
3.7
KH2PO4
1.7
AA_4
Chemical
Concentration (mg/mL)
Nicotinic acid
0.05
Pyridoxine·HCl
0.05
Thiamine·HCl
0.01
myo-Inositol
10
Store at −20°C
AA_5
Chemical
Concentration (mg/mL)
L-Glutamine
8.77
L-Aspartic acid
2.66
L-Arginine
2.28
Glycine
0.75
Store at −20°C
2,4-D (0.2 mg/mL)
Chemical
Concentration (mg/mL)
2,4-D sodium monohydrate
0.236
(2,4-Dichlorophenoxy)acetic acid sodium salt monohydrate, Sigma-Aldrich (D6679)
Kinetin (0.2 mg/mL)
Chemical
Concentration (mg/mL)
Kinetin
0.2
Dissolve kinetin in small volume of KOH (1 N), and fill up with distilled water
Agar, powder
KOH (1 N)
Medium preparation
Dissolve the following chemicals in approximately 800 mL of distilled water.
Chemical
Amount
Sucrose
30 g
Add following stock solutions, and fill up to approximately 950 mL with distilled water.
Stock solution
Volume (mL)
MS_micro_1
10
MS_micro_2
10
AA_1
100
AA_4
10
AA_5
100
2,4-D (0.2 mg/mL)
5
Kinetin (0.2 mg/mL)
1
Adjust the pH of the solution to 5.8 with KOH (1 N), and fill up to 1 L with distilled water.
Add 12 g of agar to the medium.
Autoclave the medium at 121°C for 20 min.
Formulation
Chemical
Concentration (mg/L)
L-Glutamine
877
L-Aspartic acid
266
L-Arginine
228
CaCl2·2H2O
440
MgSO4·7H2O
370
KH2PO4
170
H3BO3
6.2
MnSO4·5H2O
24.1
ZnSO4·7H2O
8.6
KI
0.83
Na2MoO4·2H2O
0.25
CuSO4·5H2O
0.025
CoCl2·6H2O
0.025
FeSO4·7H2O
27.8
Na2-EDTA
37.3
Nicotinic acid
0.5
Pyridoxine·HCl
0.5
Thiamine·HCl
0.1
Glycine
75
myo-Inositol
100
Sucrose
30000
2,4-D sodium monohydrate
1.18
Kinetin
0.2
Agar
12000