medium no. 15: AA medium, 1 mg/L 2,4-dichlorophenoxyacetic acid, 0.2 mg/L kinetin, 1.2% (w/v) agar, pH 5.8

Chemicals and stock solutions

(All stock solutions are stored at 4°C)
  • 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

  • AA_1

    Chemical Concentration (mg/mL)
    KCl 29.4
    CaCl2·2H2O 4.4
    MgSO4·7H2O 3.7
    KH2PO4 1.7
  • AA_4

    Chemical Concentration (mg/mL)
    Nicotinic acid 0.05
    Pyridoxine·HCl 0.05
    Thiamine·HCl 0.01
    myo-Inositol 10

    Store at -20°C

  • AA_5

    Chemical Concentration (mg/mL)
    L-Glutamine 8.77
    L-Aspartic acid 2.66
    L-Arginine 2.28
    Glycine 0.75

    Store at -20°C

  • 2,4-D (0.2 mg/mL)

    Chemical Concentration (mg/mL)
    2,4-D sodium monohydrate 0.236

    (2,4-Dichlorophenoxy)acetic acid sodium salt monohydrate, Sigma-Aldrich (D6679)

  • 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

  1. Dissolve the following chemicals in approximately 800 mL of distilled water.

    Chemical Amount (g)
    Sucrose 30
  2. 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
    AA_1 100
    AA_4 10
    AA_5 100
    2,4-D (0.2 mg/mL) 5
    Kinetin (0.2 mg/mL) 1
  3. Adjust the pH of the solution to 5.8 with KOH (1 N), and fill up to 1 L with distilled water.

  4. Add 12 g of agar to the medium.

  5. Autoclave the medium at 121°C for 20 min.

Formulation

Chemical Concentration (mg/L)
KCl 2940
L-Glutamine 877
L-Aspartic acid 266
L-Arginine 228
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 75
myo-Inositol 100
Sucrose 30000
2,4-D sodium monohydrate 1.18
Kinetin 0.2
Agar 12000