===============================================================================
medium no. 5: Jouanneau and Péaud-Lenoël medium, 1 µM 1-naphthaleneacetic acid
===============================================================================

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

* :doc:`/cell_lines/rpc00008`

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

(All stock solutions are stored at 4°C)

* JPL_Aʼ

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

       "KNO\ :sub:`3`", "65.5"
       "CaCl\ :sub:`2`\ ·2H\ :sub:`2`\ O", "4.4"
       "MgSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "3.7"
       "KH\ :sub:`2`\ PO\ :sub:`4`", "1.7"

* JPL_B

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

       "H\ :sub:`3`\ BO\ :sub:`3`", "6.2"
       "MnSO\ :sub:`4`\ ·5H\ :sub:`2`\ O", "24.1"
       "ZnSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "10.6"
       "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"

* JPL_C

    .. 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

* JPL_D

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

       "Glycine", "0.2"
       "*myo*-Inositol", "10"

* JPL_VT

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

       "Nicotinic acid", "0.5"
       "Pyridoxine·HCl", "0.5"
       "Thiamine·HCl", "0.4"

* JPL_P

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

       "200 mM KH\ :sub:`2`\ PO\ :sub:`4`", "19.5"
       "200 mM Na\ :sub:`2`\ HPO\ :sub:`4`", "30.5"
       "H\ :sub:`2`\ O", "50"

* NAA (1 mM)

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

       "NAA·K", "0.224"

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

* Sucrose

* Casein hydrolysate, vitamin free

    Casamino acids vitamin assay, Difco (#228820)

* KOH (1 N)

* HCl (1 N)

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

#. Prepare three solutions as follows.

  * JPL mineral solution (1 L)

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

         "JPL_Aʼ", "37.5"
         "JPL_B", "0.375"
         "JPL_C", "2.5"

      Adjust the pH to 5.7 with KOH (1 N)

  * JPL organic solution (100 mL)

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

         "JPL_D", "10 mL"
         "JPL_VT", "1 mL"
         "Casein hydrolysate", "0.1 g"

      Adjust the pH to 5.7 with HCl (1 N)

  * JPL sucrose solution (100 mL)

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

         "JPL_P", "1 mL"
         "NAA (1 mM)", "1 mL"
         "Sucrose", "15 g"

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

#. Add 800 mL of JPL mineral solution, 100 mL of JPL organic solution, and 100 mL of JPL sucrose solution asceptically.

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

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

       "KNO\ :sub:`3`", "1965"
       "CaCl\ :sub:`2`\ ·2H\ :sub:`2`\ O", "132"
       "MgSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "111"
       "KH\ :sub:`2`\ PO\ :sub:`4`", "56.308"
       "Na\ :sub:`2`\ HPO\ :sub:`4`", "8.66"
       "H\ :sub:`3`\ BO\ :sub:`3`", "1.86"
       "MnSO\ :sub:`4`\ ·5H\ :sub:`2`\ O", "7.23"
       "ZnSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "3.18"
       "KI", "0.249"
       "Na\ :sub:`2`\ MoO\ :sub:`4`\ ·2H\ :sub:`2`\ O", "0.075"
       "CuSO\ :sub:`4`\ ·5H\ :sub:`2`\ O", "0.0075"
       "CoCl\ :sub:`2`\ ·6H\ :sub:`2`\ O", "0.0075"
       "FeSO\ :sub:`4`\ ·7H\ :sub:`2`\ O", "5.56"
       "Na\ :sub:`2`\ -EDTA", "7.46"
       "Nicotinic acid", "0.5"
       "Pyridoxine·HCl", "0.5"
       "Thiamine·HCl", "0.4"
       "Glycine", "2"
       "*myo*-Inositol", "100"
       "Casein hydrolysate, vitamin free", "100"
       "Sucrose", "15000"
       "NAA·K", "0.224"