medium no. 41: modified Gamborg’s B5 medium, 0.54 mg/L 1-naphthaleneacetic acid, 0.2 mg/L kinetin, pH 5.7

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

    Potassium 1-naphthylacetate, Wako Pure Chemical Industries (# 161-04021)

  • 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

  • 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 19.85 g
  2. Add following stock solutions, and fill up to approximately 950 mL with distilled water.

    Stock solution Volume (mL)
    B5_VT 5
    NAA (1 mg/mL) 0.54
    Kinetin (0.2 mg/mL) 1
  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 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 2
Pyridoxine·HCl 2
Thiamine·HCl 20
myo-Inositol 200
Sucrose 19850
NAA·K 0.648
Kinetin 0.2