medium no. 61: Murashige and Skoog medium, 0.5 mg/L 1-naphthaleneacetic acid, 0.05 mg/L kinetin, pH 5.8

Plant cell lines

Chemicals and stock solutions

(All stock solutions are stored at 4°C)

  • MS salt mix

    Murashige and Skoog Plant Salt Mixture, FUJIFILM Wako Pure Chemical Corporation (#392-00591)

  • Sucrose

  • 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 (0.5 mg/mL)

    Chemical

    Concentration (mg/mL)

    NAA·K

    0.6

    Potassium 1-naphthylacetate, FUJIFILM Wako Pure Chemical Corporation (#161-04021); Sterilize the solution by membrane filtration, and store at −20°C

  • Kinetin (0.05 mg/mL)

    Chemical

    Concentration (mg/mL)

    Kinetin

    0.05

    Dissolve kinetin in small volume of KOH (1 N), and fill up with distilled water; Sterilize the solution by membrane filtration, and store at −20°C

  • KOH (1 N)

Medium preparation

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

    Chemical

    Amount

    MS salt mix

    1 bag (1 L)

    Sucrose

    30 g

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

    Stock solution

    Volume (mL)

    MS_VT

    1

    MS_inositol

    2.5

  3. Adjust the pH of the solution to 5.8 with KOH (1 N), and fill up to 1 L with distilled water.

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

  5. Add the following phytohormones to the medium just before subculturing.

    Chemical

    Amount

    NAA (0.5 mg/mL)

    50 µL

    Kinetin (0.05 mg/mL)

    50 µL

Formulation

Chemical

Concentration (mg/L)

KNO3

1900

NH4NO3

1650

CaCl2·2H2O

440

MgSO4·7H2O

370

KH2PO4

170

H3BO3

6.2

MnSO4·4H2O

22.3

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

30000

NAA·K

0.6

Kinetin

0.05