medium no. 32: Linsmaier and Skoog medium, 1 µM indole-3-acetic acid, 10 µM kinetin, 1% (w/v) agar, pH 6.3

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

  • IAA (1 mM)

    Chemical

    Concentration (mg/mL)

    IAA·K

    0.213

    Potassium 3-indoleacetate, FUJIFILM Wako Pure Chemical Corporation (#160-07531)

  • 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)

  • NaOH (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

    IAA (1 mM)

    1

    Kinetin (1 mM)

    10

  3. Adjust the pH of the solution to 6.3 with NaOH (1 N), and fill up to 1 L with distilled water.

  4. Add 10 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

IAA·K

0.213

Kinetin

2.15

Agar

10000