Evo m mlv reverse transcription premixed kit

The HTR-8/Svneo cells were exposed to concentrations of 0, 12.5, 25, and 37.5 μM oridonin for 24 h. As previously specified, the RNA extraction was subsequently executed, quantified to 1 μg. Then, the RNA underwent reverse transcription into cDNA using Evo M-MLV reverse transcription premixed kit (Accurate Biology, Changsha, China) for RT-qPCR. The reaction took place in a 20 μL volume, with 10 μL SYBR Green qPCR Mix, 2 μL template, and 10 μM primers (Monad, Suzhou, China). The relative gene expression under oridonin treatment was determined utilizing the 2^−ΔΔCt method [41 (link)], with the fold change compared to the control group. Table 1 displays the RT-qPCR primer sequences employed in this investigation. GAPDH was designated as the endogenous control for the remaining target genes.

Used Trizol kit (TIANGEN BIOTECH CO., LTD.) to extract total RNA from tubers of pigmented potatoes in different growth periods. Use Evo M-MLV reverse transcription premixed kit (including gDNA removal reagent for qPCR) Ver 2 (Accurate Biology), used SYBR ^® Green Pro Taq HS premixed qPCR kit (including ROX) (Accurate Biology) conducts real-time fluorescent quantitative PCR reaction. StGAPDH was the reference gene. The cDNA obtained by reverse transcription was diluted 10 times and used as a template. The primers of all genes (Table 1) were designed with SnapGene 4.3.6 software and synthesized by Sangon Biotech (Shanghai) Co., Ltd. All primers were diluted 10 times before being used. Reaction procedure: 95 °C 2 min, 95 °C 15 sec, 60 °C 30 sec, 40 cycles, Melt Curve Stage, 2^{- ΔΔ Ct} method (Livak and Schmittgen, 2001 (link)).