Quick rna isolation kit

qRT-PCR was performed to analyze the expression levels of SmePPO, SmePOD, SmeLOX, Sme PAL, SmeC4H, Sme4CL, SmeSOD, SmeCAT, SmeAPX gene. Total RNA was extracted using the Quick RNA isolation Kit (Beijing TsingKe Biotech Co., Ltd., Beijing, China) according to the manufacturer’s protocol. First-strand cDNA was synthesized from 2 μg of total RNA using M-MLV reverse transcriptase (Beijing TsingKe Biotech Co., Ltd., Beijing, China) and Oligo (dT)18 in a 25-μL reaction. Real-time PCR was performed with SYBR Green PCR mix (Takara, Shiga, Japan). SmPKG was used as an endogenous control gene for qRT-PCR analyses. Relative expression levels of the target genes were calculated using the 2− ΔΔCt method. The primers used are listed in Table S3.

qRT-PCR was performed to verify the accuracy of the gene expression profile obtained from the RNA-seq data. Total RNA was extracted using the Quick RNA isolation Kit (Beijing Tsingke Biotech, Beijing, China) according to the manufacturer’s protocol. First-strand cDNA was synthesized from 2 μg total RNA using M-MLV reverse transcriptase (Beijing Tsingke Biotech) and Oligo (dT)18 in a 25 μL reaction. Real-time PCR was performed with SYBR Green PCR mix (TaKaRa, Shiga, Japan). SmPKG was used as an endogenous control gene for qRT-PCR analyses. Relative expression levels of the target genes were calculated using the 2^−ΔΔCt method. The primers used are listed in Supplementary Table S5.