medium no. 2: modified Lin and Staba medium, 0.5 µM 2,4-dichlorophenoxyacetic acid, pH 5.7
Plant cell lines
Chemicals and stock solutions
(All stock solutions are stored at 4°C)
KNO3
NH4Cl
CaCl2·2H2O
MgSO4·7H2O
Sucrose
Lin_and_Staba_1
Chemical
Concentration (mg/mL)
H3BO3
2.4
MnSO4·5H2O
10.18
ZnSO4·7H2O
4.05
Lin_and_Staba_2
Chemical
Concentration (mg/mL)
FeSO4·7H2O
5.56
Na2-EDTA
7.46
Heat at 80°C for 3–4 hours for chelating Fe
Lin_and_Staba_3
Chemical
Concentration (mg/mL)
KI
0.375
Na2MoO4·2H2O
0.127
CuSO4·5H2O
0.01
CoCl2·6H2O
0.01
Lin_and_Staba_4
Chemical
Concentration (mg/mL)
Nicotinic acid
5
Pyridoxine·HCl
0.5
Thiamine·HCl
3
Lin_and_Staba_5
Chemical
Concentration (mg/mL)
KH2PO4
68
2,4-D (1 mM)
Chemical
Concentration (mg/mL)
2,4-D sodium monohydrate
0.261
(2,4-Dichlorophenoxy)acetic acid sodium salt monohydrate, Sigma-Aldrich (D6679)
KOH (1 N)
Medium preparation
Dissolve the following chemicals in approximately 800 mL of distilled water.
Chemical
Amount
KNO3
5.56 g
NH4Cl
0.268 g
CaCl2·2H2O
0.22 g
MgSO4·7H2O
0.185 g
Sucrose
20 g
Add following stock solutions, and fill up to approximately 950 mL with distilled water.
Stock solution
Volume (mL)
Lin_and_Staba_1
1
Lin_and_Staba_2
5
Lin_and_Staba_3
1
Lin_and_Staba_4
1
Lin_and_Staba_5
1
2,4-D (1 mM)
0.5
Adjust the pH of the solution to 5.7 with KOH (1 N), and fill up to 1 L with distilled water.
Autoclave the medium at 121°C for 20 min.
Formulation
Chemical
Concentration (mg/L)
KNO3
5560
NH4Cl
268
CaCl2·2H2O
220
MgSO4·7H2O
185
KH2PO4
68
H3BO3
2.4
MnSO4·5H2O
10.18
ZnSO4·7H2O
4.05
KI
0.375
Na2MoO4·2H2O
0.127
CuSO4·5H2O
0.01
CoCl2·6H2O
0.01
FeSO4·7H2O
27.8
Na2-EDTA
37.3
Nicotinic acid
5
Pyridoxine·HCl
0.5
Thiamine·HCl
3
Sucrose
20000
2,4-D sodium monohydrate
0.131