Solanum lycopersicum (tomato) cell lines

Plant cell lines

Genotypes

Cell line

rbcL

matK

Tm-1

Sly-1

Solanum lycopersicum

Sly-1

GCR26

Solanum lycopersicum

GCR

+/+

GCR237

Solanum lycopersicum

GCR

Tm-1/Tm-1

Protocols

DNA barcoding — rbcL

DNA barcoding — matK

  1. PCR

    • Primer set 《Primer sequences

      Target

      Forward primer

      Reverse primer

      Product size

      matK

      matK_F

      matK_R

      900 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

      Template DNA

      1 µL

      Sterile distilled water

      3.75 µL

    • PCR reaction

      Temperature

      Time

      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 sequence of the PCR product.

    • NCBI BLAST search

    • Compare the DNA sequence with that of previous years.

DNA sequencing — Tm-1 gene

  1. PCR

    • Primer set 《Primer sequences

      Target

      Forward primer

      Reverse primer

      PCR product size

      Tm-1

      GCR_F2

      GCRseq_R1

      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

      Template DNA

      1 µL

      Sterile distilled water

      3.75 µL

    • PCR reaction

      Temperature

      Time

      Cycles

      94°C

      1 min

      94°C

      30 sec

      35 cycles

      55°C

      30 sec

      72°C

      1 min

      72°C

      7 min

  2. Check the DNA sequence of the PCR product.