medium no. 33: modified Linsmaier and Skoog medium, 1 µM indole-3-acetic acid, 10 µM kinetin, 1% (w/v) agar, pH 6.3
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 Corporation (#392-00591)
Sucrose
CuSO4 (1.2 mM)
Chemical
Concentration (mg/mL)
CuSO4·5H2O
0.3
LS_VT
Chemical
Concentration (mg/mL)
Thiamine·HCl
0.16
myo-Inositol
40
IAA (1 mM)
Chemical
Concentration (mg/mL)
IAA·K
0.213
Potassium 3-indoleacetate, FUJIFILM Wako Pure Chemical Corporation (# 160-07531)
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
NaOH (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)
CuSO4 (1.2 mM)
1
LS_VT
2.5
IAA (1 mM)
1
Kinetin (1 mM)
10
Adjust the pH of the solution to 6.3 with NaOH (1 N), and fill up to 1 L with distilled water.
Add 10 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
0.325
CoCl2·6H2O
0.025
FeSO4·7H2O
27.8
Na2-EDTA
37.3
Thiamine·HCl
0.4
myo-Inositol
100
Sucrose
30000
IAA·K
0.213
Kinetin
2.15
Agar
10000