Primescript 4 1st strand cdna synthesis kit

cDNA was synthesized from 500-ng aliquots of total RNA from Japanese cedar and A. thaliana using the PrimeScript IV 1st Strand cDNA synthesis kit (Takara Bio, Shiga, Japan). CjTKPR1 and EF1-α were amplified using intron-containing primers to distinguish between cDNA and gDNA amplification. The primer sequences are listed in Table S5.

cDNA was synthesized from 500-ng aliquots of total RNA from Japanese cedar and A. thaliana using the PrimeScript IV 1st Strand cDNA synthesis kit (Takara Bio, Shiga, Japan). CjTKPR1 and EF1-α were amplified using intron-containing primers to distinguish between cDNA and gDNA amplification. The primer sequences are listed in Table S5.

Treated cells were harvested with RNAios Plus (Takara #9109, Japan) and extracted with an equal volume of chloroform. After centrifugation at 4 °C and 12,000 × g for 15 min, the top supernatant layer was carefully harvested and mixed with the same volume of isopropanol. After 2 h freezing, the samples were then centrifuged at 4 °C and 12000 × g for 10 min. The precipitates were then rinsed with 75% ethanol in DEPC water (Solarbio R1600, China) and air dried. The precipitated RNA was then dissolved in DEPC water (Solarbio R1600, China) and quantified with a Nanodrop (Thermo Scientific NanoDrop 8000, USA). Reverse transcription was performed using the PrimeScript IV 1st strand cDNA synthesis kit (TaKaRa #6215, Japan) according to the standard protocol.