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

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

* :doc:`/cell_lines/index`

Genotypes
=========

* :doc:`../../genotyping/index`

Protocol
========

#. PCR

   * Primer sets 《:doc:`_primers`》

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

          "rbcL (long)", "13", "18", "1.4 kbp"
          "rbcL (partial)", "13", "rbcL-R", "600 bp"

   * PCR reaction mixture

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

          "TaKaRa Ex Taq™ Premix", "5 µL"
          "Forward primer (100 µM)", "0.125 µL"
          "Reverse primer (100 µM)", "0.125 µL"
          "Genome DNA", "1 µL"
          "Sterile distilled water", "3.75 µL"

   * PCR reaction

       +-----------+--------+---------+
       |Temperature|Duration|Cycles   |
       +===========+========+=========+
       |94°C       |1 min   |         |
       +-----------+--------+---------+
       |94°C       |30 sec  |35 cycles|
       +-----------+--------+         +
       |55°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.