medium no. 48: Gamborg’s B5 medium, 1 mg/L 1-naphthaleneacetic acid, 0.01 mg/L kinetin, 0.8% agar, pH 5.8

Materials

Chemicals and stock solutions

(All stock solutions are stored at 4°C)
  • B5 salt mix

    Gamborg’s B5 Medium Salt Mixture, Wako Pure Chemical Industries (#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

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

    Chemical Amount
    B5 salt mix 1 bag (1 L)
    Sucrose 20 g
  2. 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
  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 8 g of agar to the medium.

  5. 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