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medium no. 43: modified Murashige and Skoog medium, 1 mg/L 2,4-dichlorophenoxyacetic acid, 0.8% (w/v) agar, pH 5.8
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Plant cell lines
=================

* :doc:`/cell_lines/rpc00055`

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

* KH\ :sub:`2`\ PO\ :sub:`4` (100 mg/mL)

    .. csv-table::
       :header: "Chemical", "Concentration (mg/mL)"
       :widths: auto

       "KH\ :sub:`2`\ PO\ :sub:`4`", "100"

* B5_VT

    .. csv-table::
       :header: "Chemical", "Concentration (mg/mL)"
       :widths: auto

       "Nicotinic acid", "0.4"
       "Pyridoxine·HCl", "0.4"
       "Thiamine·HCl", "4"
       "*myo*-Inositol", "40"

* 2,4-D (0.2 mg/mL)

    .. csv-table::
       :header: "Chemical", "Concentration (mg/mL)"
       :widths: auto

       "2,4-D sodium monohydrate", "0.236"

    (2,4-Dichlorophenoxy)acetic acid sodium salt monohydrate, Sigma-Aldrich (D6679)

* Agar, powder

* KOH (1 N)

Medium preparation
===================

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

    .. csv-table::
       :header: "Chemical", "Amount"
       :widths: auto

       "MS salt mix", "1 bag (1 L)"
       "Sucrose", "20 g"

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

    .. csv-table::
       :header: "Stock solution", "Volume (mL)"
       :widths: auto

       "KH\ :sub:`2`\ PO\ :sub:`4` (100 mg/mL)", "3.4"
       "B5_VT", "5"
       "2,4-D (0.2 mg/mL)", "5"

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

#. Add 8 g of agar to the medium.

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

Formulation
============

    .. csv-table::
       :header: "Chemical", "Concentration (mg/L)"
       :widths: auto

       "KNO\ :sub:`3`", "1900"
       "NH\ :sub:`4`\ NO\ :sub:`3`", "1650"
       "CaCl\ :sub:`2`\ ·2H\ :sub:`2`\ O", "440"
       "MgSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "370"
       "KH\ :sub:`2`\ PO\ :sub:`4`", "510"
       "H\ :sub:`3`\ BO\ :sub:`3`", "6.2"
       "MnSO\ :sub:`4`\ ·4H\ :sub:`2`\ O", "22.3"
       "ZnSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "8.6"
       "KI", "0.83"
       "Na\ :sub:`2`\ MoO\ :sub:`4`\ ·2H\ :sub:`2`\ O", "0.25"
       "CuSO\ :sub:`4`\ ·5H\ :sub:`2`\ O", "0.025"
       "CoCl\ :sub:`2`\ ·6H\ :sub:`2`\ O", "0.025"
       "FeSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "27.8"
       "Na\ :sub:`2`\ -EDTA", "37.3"
       "Nicotinic acid", "2"
       "Pyridoxine·HCl", "2"
       "Thiamine·HCl", "20"
       "*myo*-Inositol", "200"
       "Sucrose", "20000"
       "2,4-D sodium monohydrate", "1.18"
       "Agar", "8000"