Transgene insertion

Plant cell lines

Genotypes

Cell line

Transgene insertion

CRA

Not detected (tmr)

V208

tmr

GV7

35Spro-GFP-AtVam3p

GF11

35Spro-GFP-AtFim1

GT16

35Spro-GFP-tubulin

TBY2-AtRER1B

35Spro-GFP-AtRer1B

BY-TIPG

35Spro-NtTIP1;1-GFP

156-S

35Spro-CjSMT

TBY2-31/ST

35Spro-GFP-AtSYP31/35Spro-ST-mRFP

TBY2-41/ST

35Spro-GFP-AtSYP41/35Spro-ST-mRFP

TBY2-R31

35Spro-mRFP-AtSYP31

Protocol

  1. PCR

    • Primer sets 《Primer sequences

      Cell line

      Target

      Forward

      Reverse

      Product size

      CRA

      tmr

      35Spro_F3

      GV7_R

      Not detected (370 bp)

      V208

      tmr

      35Spro_F3

      GV7_R

      370 bp

      GV7

      35Spro-GFP-AtVam3p

      35Spro_F3

      GV7_R

      1.5 kbp

      GV7

      35Spro-GFP-AtVam3p

      GFP_F

      GV7_R

      1.3 kbp

      GF11

      35Spro-GFP-AtFim1

      35Spro_F3

      GF11_R

      2.0 kbp

      GF11

      35Spro-GFP-AtFim1

      GFP_F

      GF11_R

      1.7 kbp

      GT16

      35Spro-GFP-tubulin

      35Spro_F3

      GT16_R

      2.1 kbp

      GT16

      35Spro-GFP-tubulin

      GFP_F

      GT16_R

      1.9 kbp

      TBY2-AtRER1B

      35Spro-GFP-AtRer1B

      35Spro_F3

      RER1B_R

      1.4 kbp

      TBY2-AtRER1B

      35Spro-GFP-AtRer1B

      GFP_494_F

      RER1B_R

      800 bp

      TBY2-AtRER1B

      35Spro-GFP-AtRer1B

      GFP_F

      RER1B_R

      1.2 kbp

      TBY2-AtRER1B

      35Spro-GFP-AtRer1B

      GFPS65T_F

      RER1B_R1

      565 bp

      BY-TIPG

      35Spro-NtTIP1;1-GFP

      35Spro_F3

      GFP_R

      1.3 kbp

      BY-TIPG

      35Spro-NtTIP1;1-GFP

      NtTIP1_F

      GFP_R

      1.0 kbp

      156-S

      35Spro-CjSMT

      35Spro_F2

      SMT_R

      1.5 kbp

      156-S

      35Spro-CjSMT

      35Spro_F2

      NOSterR1

      2.0 kbp

      TBY2-31/ST

      35Spro-GFP-AtSYP31

      35Spro_F5

      SYP31stop_R

      1.8 kbp

      TBY2-31/ST

      35Spro-ST-mRFP

      35Spro_F2

      mRFPstop_R

      1.1 kbp

      TBY2-41/ST

      35Spro-GFP-AtSYP41

      35Spro_F5

      SYP41stop_R

      1.8 kbp

      TBY2-41/ST

      35Spro-ST-mRFP

      35Spro_F2

      mRFPstop_R

      1.1 kbp

      TBY2-R31

      35Spro-mRFP-AtSYP31

      35Spro_F5

      SYP31stop_R

      1.8 kbp

    • PCR reaction mixture

      Component

      10 µL reaction

      TaKaRa Ex Taq™ Premix

      5 µL

      Forward primer (10 µM)

      0.5 µL

      Reverse primer (10 µM)

      0.5 µL

      Template DNA

      1 µL

      Sterile distilled water

      3 µL

    • PCR reaction

      Temperature

      Time

      Cycles

      94°C

      1 min

      94°C

      30 sec

      35 cycles

      55°C

      30 sec

      72°C

      1 min/1 kbp

      72°C

      10 min

  2. Check the size of the PCR product.

  3. If necessary, check the DNA sequence of the PCR product.