DNA barcoding — rbcL

Plant cell lines

Genotypes

Protocol

  1. PCR

    • Primer sets 《Primer sequences》

      Target

      Forward primer

      Reverse primer

      PCR product size

      rbcL (long)

      13

      18

      1.4 kbp

      rbcL (partial)

      13

      rbcL-R

      600 bp

    • PCR reaction mixture

      Component

      10 µL reaction

      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

  2. Check the DNA sequence of the PCR product.

    • NCBI BLAST search

    • Compare the DNA sequence with that of previous years.