medium no. 2: modified Lin and Staba medium, 0.5 µM 2,4-dichlorophenoxyacetic acid, pH 5.7

Chemicals and stock solutions

(All stock solutions are stored at 4°C)
  • KNO3

  • NH4Cl

  • CaCl2·2H2O

  • MgSO4·7H2O

  • Sucrose

  • Lin_and_Staba_1

    Chemical Concentration (mg/mL)
    H3BO3 2.4
    MnSO4·5H2O 10.18
    ZnSO4·7H2O 4.05
  • Lin_and_Staba_2

    Chemical Concentration (mg/mL)
    FeSO4·7H2O 5.56
    Na2-EDTA 7.46

    Heat at 80°C for 3–4 hours for chelating Fe

  • Lin_and_Staba_3

    Chemical Concentration (mg/mL)
    KI 0.375
    Na2MoO4·2H2O 0.127
    CuSO4·5H2O 0.01
    CoCl2·6H2O 0.01
  • Lin_and_Staba_4

    Chemical Concentration (mg/mL)
    Nicotinic acid 5
    Pyridoxine·HCl 0.5
    Thiamine·HCl 3
  • Lin_and_Staba_5

    Chemical Concentration (mg/mL)
    KH2PO4 68
  • 2,4-D (1 mM)

    Chemical Concentration (mg/mL)
    2,4-D sodium monohydrate 0.261

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

  • KOH (1 N)

Medium preparation

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

    Chemical Amount (g)
    KNO3 5.56
    NH4Cl 0.268
    CaCl2·2H2O 0.22
    MgSO4·7H2O 0.185
    Sucrose 20
  2. Add following stock solutions, and fill up to approximately 950 mL with distilled water.

    Stock solution Volume (mL)
    Lin_and_Staba_1 1
    Lin_and_Staba_2 5
    Lin_and_Staba_3 1
    Lin_and_Staba_4 1
    Lin_and_Staba_5 1
    2,4-D (1 mM) 0.5
  3. Adjust the pH of the solution to 5.7 with KOH (1 N), and fill up to 1 L with distilled water.

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

Formulation

Chemical Concentration (mg/L)
KNO3 5560
NH4Cl 268
CaCl2·2H2O 220
MgSO4·7H2O 185
KH2PO4 68
H3BO3 2.4
MnSO4·5H2O 10.18
ZnSO4·7H2O 4.05
KI 0.375
Na2MoO4·2H2O 0.127
CuSO4·5H2O 0.01
CoCl2·6H2O 0.01
FeSO4·7H2O 27.8
Na2-EDTA 37.3
Nicotinic acid 5
Pyridoxine·HCl 0.5
Thiamine·HCl 3
Sucrose 20000
2,4-D sodium monohydrate 0.131