medium no. 13: modified Murashige and Skoog medium, 0.5 mg/L 1-naphthaleneacetic acid, 1 mg/L 6-benzylaminopurine, pH 6.5

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, Wako Pure Chemical Industries (# 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

  • NaOH (1 N)

Medium preparation

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

    Chemical Amount (g)
    KCl 1.4
    L-Glutamine 1
    NZ-Amine Type A 1
    CaCl2·2H2O 0.44
    MgSO4·7H2O 0.37
    KH2PO4 0.17
    Sucrose 20
  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
    MS_VT 1
    MS_inositol 2.5
    NAA (1 mg/mL) 0.5
    BAP (1 mg/mL) 1
  3. Adjust the pH of the solution to 6.5 with NaOH (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)
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