medium no. 8: Linsmaier and Skoog medium, 0.1 µM 2,4-dichlorophenoxyacetic acid, 1.2% (w/v) agar, pH 5.7
Plant cell lines
Chemicals and stock solutions
(All stock solutions are stored at 4°C)
MS salt mix
Murashige and Skoog Plant Salt Mixture, FUJIFILM Wako Pure Chemical Corporations (#392-00591)
Sucrose
LS_VT
Chemical
Concentration (mg/mL)
Thiamine·HCl
0.16
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)
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
MS salt mix
1 bag (1 L)
Sucrose
30 g
Add following stock solutions, and fill up to approximately 950 mL with distilled water.
Stock solution
Volume (mL)
LS_VT
2.5
2,4-D (1 mM)
0.1
Adjust the pH of the solution to 5.7 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)
KNO3
1900
NH4NO3
1650
CaCl2·2H2O
440
MgSO4·7H2O
370
KH2PO4
170
H3BO3
6.2
MnSO4·4H2O
22.3
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
Thiamine·HCl
0.4
myo-Inositol
100
Sucrose
30000
2,4-D sodium monohydrate
0.0261
Agar
12000