medium no. 22: Linsmaier and Skoog medium, 1 µM 1-naphthaleneacetic acid, 1 µM kinetin, 1.2% (w/v) agar, pH 5.6

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

  • LS_VT

    Chemical

    Concentration (mg/mL)

    Thiamine·HCl

    0.16

    myo-Inositol

    40

  • NAA (1 mM)

    Chemical

    Concentration (mg/mL)

    NAA·K

    0.224

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

  • Kinetin (1 mM)

    Chemical

    Concentration (mg/mL)

    Kinetin

    0.215

    Dissolve kinetin in small volume of KOH (1 N), and fill up with distilled water

  • Agar, powder

    Agar, powder, Junsei Chemical (#24440-1201)

  • 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)

    LS_VT

    2.5

    NAA (1 mM)

    1

    Kinetin (1 mM)

    1

  3. Adjust the pH of the solution to 5.6 with KOH (1 N), and fill up to 1 L with distilled water.

  4. Add 12 g of agar to the medium.

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

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

Thiamine·HCl

0.4

myo-Inositol

100

Sucrose

30000

NAA·K

0.224

Kinetin

0.215

Agar

12000