medium no. 68: modified Murashige and Skoog medium, 0.5 mg/L 1-naphthaleneacetic acid, 1 mg/L 6-benzylaminopurine, 1.2% (w/v) agar, pH 6.5
Plant cell lines
Chemicals and stock solutions
(All stock solutions are stored at 4°C)
KCl
L-Glutamine
NZ-Amine Type A
CaCl2·2H2O
MgSO4·7H2O
KH2PO4
Sucrose
MS_micro_1
Chemical
Concentration (mg/mL)
H3BO3
0.62
MnSO4·5H2O
2.41
ZnSO4·7H2O
0.86
KI
0.083
Na2MoO4·2H2O
0.025
CuSO4·5H2O
0.0025
CoCl2·6H2O
0.0025
MS_micro_2
Chemical
Concentration (mg/mL)
FeSO4·7H2O
2.78
Na2-EDTA
3.73
Heat at 80°C for 3–4 hours for chelating Fe
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
NAA (1 mg/mL)
Chemical
Concentration (mg/mL)
NAA·K
1.2
Potassium 1-naphthylacetate, FUJIFILM Wako Pure Chemical Corporation (#161-04021)
BAP (1 mg/mL)
Chemical
Concentration (mg/mL)
6-Benzylaminopurine
1
Dissolve 6-benzylaminopurine in small volume of KOH (1 N), and fill up with distilled water
Agar, powder
Agar, powder, Junsei Chemical (#24440-1201)
NaOH (1 N)
Medium preparation
Dissolve the following chemicals in approximately 800 mL of distilled water.
Chemical
Amount
KCl
1.4 g
L-Glutamine
1 g
NZ-Amine Type A
1 g
CaCl2·2H2O
0.44 g
MgSO4·7H2O
0.37 g
KH2PO4
0.17 g
Sucrose
20 g
Add following stock solutions, and fill up to approximately 950 mL with distilled water.
Stock solution
Volume (mL)
MS_micro_1
10
MS_micro_2
10
MS_VT
1
MS_inositol
2.5
NAA (1 mg/mL)
0.5
BAP (1 mg/mL)
1
Adjust the pH of the solution to 6.5 with NaOH (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)
KCl
1400
L-Glutamine
1000
NZ-Amine Type A
1000
CaCl2·2H2O
440
MgSO4·7H2O
370
KH2PO4
170
H3BO3
6.2
MnSO4·5H2O
24.1
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
20000
NAA·K
0.6
6-Benzylaminopurine
1
Agar
12000