=============================
*Lotus japonicus* cell lines
=============================

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

* :doc:`/cell_lines/rpc00032`

* :doc:`/cell_lines/rpc00033`

* :doc:`/cell_lines/rpc00034`

Genotypes
=========

  .. csv-table::
     :header: "Cell line", "Culture type", "rbcL", "atpF--atpH"
     :widths: auto

     "\ :doc:`Lj </genotyping/g_rpc00032>`\ ", "Liquid", "*Lotus japonicus*", "Lj/LjA"
     "\ :doc:`LjA </genotyping/g_rpc00033>`\ ", "Agar", "*Lotus japonicus*", "Lj/LjA"
     "\ :doc:`LjmA </genotyping/g_rpc00034>`\ ", "Agar", "*Lotus japonicus*", "LjmA"

Protocols
=========

DNA barcoding --- rbcL
----------------------

  :doc:`_rbcL`

DNA barcoding --- atpF--atpH
----------------------------

#. PCR

   * Primer set 《:doc:`_primers`》

       .. csv-table::
          :header: "Target", "Forward primer", "Reverse primer", "Product size"
          :widths: auto

          "atpF--atpH", "atpF_f", "atpH_r", "800 bp"

   * PCR reaction mixture

       .. csv-table::
          :header: "Component", "10 µL reaction"
          :widths: auto

          "TaKaRa Ex Taq™ Premix", "5 µL"
          "Forward primer (100 µM)", "0.5 µL"
          "Reverse primer (100 µM)", "0.5 µL"
          "Template DNA", "1 µL"
          "Sterile distilled water", "3 µL"

   * PCR reaction

       +-----------+------+---------+
       |Temperature|Time  |Cycles   |
       +===========+======+=========+
       |95°C       |1 min |         |
       +-----------+------+---------+
       |94°C       |30 sec|35 cycles|
       +-----------+------+         +
       |52°C       |30 sec|         |
       +-----------+------+         +
       |72°C       |1 min |         |
       +-----------+------+---------+
       |72°C       |10 min|         |
       +-----------+------+---------+

#. Check the DNA sequence of the PCR product.

   * NCBI BLAST search

   * Compare the DNA sequence with that of previous years.