====================================
*Phyllostachys* (bamboo) cell lines
====================================

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

* :doc:`/cell_lines/rpc00047`

* :doc:`/cell_lines/rpc00048`

Genotypes
=========

  .. csv-table::
     :header: "Cell line", "rbcL", "psbK--psbI"
     :widths: auto

     "\ :doc:`Pn </genotyping/g_rpc00047>`\ ", "*Phyllostachys nigra*", "*Phyllostachys nigra*"
     "\ :doc:`Pb </genotyping/g_rpc00048>`\ ", "Not detected", "*Phyllostachys bambusoides*"

Protocols
=========

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

  :doc:`_rbcL`

DNA barcoding --- psbK--psbI
----------------------------

#. PCR

   * Primer set 《:doc:`_primers`》

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

          "psbK--psbI", "psbK_f", "psbI_r", "500 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.