medium no. 49: Murashige and Skoog medium, 1 mg/L indole-3-acetic acid, 0.1 mg/L kinetin, 0.9% (w/v) agar, pH 5.8
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
MS_VT
Chemical
Concentration (mg/mL)
Nicotinic acid
0.5
Pyridoxine·HCl
0.5
Thiamine·HCl
0.1
Glycine
2
MS_inositol
Chemical
Concentration (mg/mL)
myo-Inositol
40
IAA (1 mg/mL)
Chemical
Concentration (mg/mL)
IAA·K
1.217
Potassium 3-indoleacetate, FUJIFILM Wako Pure Chemical Corporation (# 160-07531)
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
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)
MS_VT
1
MS_inositol
2.5
IAA (1 mg/mL)
1
Kinetin (0.2 mg/mL)
0.5
Adjust the pH of the solution to 5.8 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
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
Nicotinic acid
0.5
Pyridoxine·HCl
0.5
Thiamine·HCl
0.1
Glycine
2
myo-Inositol
100
Sucrose
30000
IAA·K
1.217
Kinetin
0.1
Agar
9000