Spinacia oleracea (spinach) cell lines

Plant cell lines

• rpc00016: Spinacia oleracea Spi-WT cell suspension culture

• rpc00017: Spinacia oleracea Spi-I-1 callus culture

• rpc00018: Spinacia oleracea Spi-12F cell suspension culture

Genotypes

Cell line	rbcL	Marker 8	NR intron
Spi-WT	Spinacia oleracea	Spi-WT/Spi-12F	Spi-WT
Spi-I-1	Spinacia oleracea	Spi-I-1	Spi-I-1/Spi-12F
Spi-12F	Spinacia oleracea	Spi-WT/Spi-12F	Spi-I-1/Spi-12F

Protocols

DNA barcoding — rbcL

DNA barcoding — rbcL

DNA sequencing

1. PCR

   • Primer set 《Primer sequences》

     Target	Forward primer	Reverse primer	Product size
     Marker 8	Spi8_F	Spi8_R	600 bp
     NR intron	Sp_NR_F6	Sp_NR_R1	800 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.