==========================================================================================================================
medium no. 13: modified Murashige and Skoog medium, 0.5 mg/L 1-naphthaleneacetic acid, 1 mg/L 6-benzylaminopurine, pH 6.5
==========================================================================================================================

Plant cell lines
=================

* :doc:`/cell_lines/rpc00018`

Chemicals and stock solutions
==============================

(All stock solutions are stored at 4°C)

* KCl

* L-Glutamine

* NZ-Amine Type A

* CaCl\ :sub:`2`\ ·2H\ :sub:`2`\ O

* MgSO\ :sub:`4`\ ·7H\ :sub:`2`\ O

* KH\ :sub:`2`\ PO\ :sub:`4`\

* Sucrose

* MS_micro_1

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

       "H\ :sub:`3`\ BO\ :sub:`3`", "0.62"
       "MnSO\ :sub:`4`\ ·5H\ :sub:`2`\ O", "2.41"
       "ZnSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "0.86"
       "KI", "0.083"
       "Na\ :sub:`2`\ MoO\ :sub:`4`\ ·2H\ :sub:`2`\ O", "0.025"
       "CuSO\ :sub:`4`\ ·5H\ :sub:`2`\ O", "0.0025"
       "CoCl\ :sub:`2`\ ·6H\ :sub:`2`\ O", "0.0025"

* MS_micro_2

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

       "FeSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "2.78"
       "Na\ :sub:`2`\ -EDTA", "3.73"

    Heat at 80°C for 3--4 hours for chelating Fe

* MS_VT

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

       "Nicotinic acid", "0.5"
       "Pyridoxine·HCl", "0.5"
       "Thiamine·HCl", "0.1"
       "Glycine", "2"

* MS_inositol

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

       "*myo*-Inositol", "40"

* NAA (1 mg/mL)

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

       "NAA·K", "1.2"

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

* BAP (1 mg/mL)

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

       "6-Benzylaminopurine", "1"

    Dissolve 6-benzylaminopurine in small volume of KOH (1 N), and fill up with distilled water

* NaOH (1 N)

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

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

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

       "KCl", "1.4 g"
       "L-Glutamine", "1 g"
       "NZ-Amine Type A", "1 g"
       "CaCl\ :sub:`2`\ ·2H\ :sub:`2`\ O", "0.44 g"
       "MgSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "0.37 g"
       "KH\ :sub:`2`\ PO\ :sub:`4`", "0.17 g"
       "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

       "MS_micro_1", "10"
       "MS_micro_2", "10"
       "MS_VT", "1"
       "MS_inositol", "2.5"
       "NAA (1 mg/mL)", "0.5"
       "BAP (1 mg/mL)", "1"

#. Adjust the pH of the solution to 6.5 with NaOH (1 N), and fill up to 1 L with distilled water.

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

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

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

       "KCl", "1400"
       "L-Glutamine", "1000"
       "NZ-Amine Type A", "1000"
       "CaCl\ :sub:`2`\ ·2H\ :sub:`2`\ O", "440"
       "MgSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "370"
       "KH\ :sub:`2`\ PO\ :sub:`4`", "170"
       "H\ :sub:`3`\ BO\ :sub:`3`", "6.2"
       "MnSO\ :sub:`4`\ ·5H\ :sub:`2`\ O", "24.1"
       "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", "0.5"
       "Pyridoxine·HCl", "0.5"
       "Thiamine·HCl", "0.1"
       "Glycine", "2"
       "*myo*-Inositol", "100"
       "Sucrose", "20000"
       "NAA·K", "0.6"
       "6-Benzylaminopurine", "1"