medium no. 48: Gamborgʼs B5 medium, 1 mg/L 1-naphthaleneacetic acid, 0.01 mg/L kinetin, 0.8% (w/v) agar, pH 5.8
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
NAA (1 mg/mL)
Chemical
Concentration (mg/mL)
NAA·K
1.2
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
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
NAA (1 mg/mL)
1
Kinetin (0.2 mg/mL)
0.05
Adjust the pH of the solution to 5.8 with KOH (1 N), and fill up to 1 L with distilled water.
Add 8 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
NAA·K
1.2
Kinetin
0.01
Agar
8000