medium no. 30: Gamborgʼs B5 medium, 1 µM 2,4-dichlorophenoxyacetic acid, 0.04 µM kinetin, 0.9% (w/v) agar, pH 5.7
Plant cell lines
Chemicals and stock solutions
(All stock solutions are stored at 4°C)
B5 salt mix
Gamborgʼs B5 Medium Salt Mixture, FUJIFILM Wako Pure Chemical Corporation (#399-00621)
Sucrose
B5_VT
Chemical
Concentration (mg/mL)
Nicotinic acid
0.4
Pyridoxine·HCl
0.4
Thiamine·HCl
4
myo-Inositol
40
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)
Kinetin (1 mM)
Chemical
Concentration (mg/mL)
Kinetin
0.215
Dissolve kinetin in small volume of KOH (1 N), and fill up with distilled water
Agar, powder
Agar, powder, Junsei Chemical (#24440-1201)
KOH (1 N)
Medium preparation
Dissolve the following chemicals in approximately 800 mL of distilled water.
Chemical
Amount
B5 salt mix
1 bag (1 L)
Sucrose
20 g
Add following stock solutions, and fill up to approximately 950 mL with distilled water.
Stock solution
Volume (mL)
B5_VT
2.5
2,4-D (1 mM)
1
Kinetin (1 mM)
0.04
Adjust the pH of the solution to 5.7 with KOH (1 N), and fill up to 1 L with distilled water.
Add 9 g of agar to the medium.
Autoclave the medium at 121°C for 20 min.
Formulation
Chemical
Concentration (mg/L)
KNO3
2500
(NH4)2SO4
134
CaCl2·2H2O
150
MgSO4·7H2O
250
NaH2PO4·H2O
150
H3BO3
3
MnSO4·H2O
10
ZnSO4·7H2O
2
KI
0.75
Na2MoO4·2H2O
0.25
CuSO4·5H2O
0.025
CoCl2·6H2O
0.025
FeSO4·7H2O
27.8
Na2-EDTA
37.3
Nicotinic acid
1
Pyridoxine·HCl
1
Thiamine·HCl
10
myo-Inositol
100
Sucrose
20000
2,4-D sodium monohydrate
0.261
Kinetin
0.0086
Agar
8000