Lotus japonicus cell lines

Plant cell lines

Genotypes

Cell line

Culture type

rbcL

atpF–atpH

Lj

Liquid

Lotus japonicus

Lj/LjA

LjA

Agar

Lotus japonicus

Lj/LjA

LjmA

Agar

Lotus japonicus

LjmA

Protocols

DNA barcoding — rbcL

DNA barcoding — atpF–atpH

  1. PCR

    • Primer set 《Primer sequences

      Target

      Forward primer

      Reverse primer

      Product size

      atpF–atpH

      atpF_f

      atpH_r

      800 bp

    • PCR reaction mixture

      Component

      10 µL reaction

      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

  2. Check the DNA sequence of the PCR product.

    • NCBI BLAST search

    • Compare the DNA sequence with that of previous years.