Tissue total rna isolation kit

Total RNA was extracted using Tissue Total RNA Isolation Kit (Vazyme, Nanjing, China), followed by a reverse transcription using HiScript II Reverse Transcriptase (Vazyme, Nanjing, China). SYBR green qPCR Mix (Vazyme, Nanjing, China) was used for the qRT-PCR experiment on the LightCycler^® 96 real-time PCR assay system (Roche, Switzerland). SmUbiquitin served as the reference gene.

Total RNA was extracted using the Tissue Total RNA Isolation Kit (Vazyme, Nanjing, China) and reverse transcribed to cDNA using HiScript II Reverse Transcriptase (Vazyme, Nanjing, China). The qRT-PCR was performed using the SYBR green qPCR Mix (Vazyme, Nanjing, China) using a real-time fluorescence quantitative PCR detection system (Roche). SmUbiquitin served as an internal control. The expression levels of SmSPL6 and other genes were calculated by the 2^−ΔΔCT analysis method [54 (link)].

Total hepatic RNA from mouse liver tissues was extracted using a tissue total RNA isolation kit according to the manufacturer’s instruction (Vazyme, CHN). Purification of 500 ng RNA and reverse transcription to cDNA by reverse transcription PCR was performed using a HiScript II Q RT SuperMix for qPCR (Vazyme, CHN). Quantitative real-time PCR was performed using a ChamQ Universal SYBR qPCR Master Mix (Vazyme, CHN). The reaction conditions were as follows: 95°C for 30 s, followed by 40 cycles of denaturation at 95°C for 10 s and extension at 60°C for 30 s. The primers were shown in Table 1. The results were analyzed using the QuantStudio 7 Flex detection system (Applied Biosystems Co., United States) and the 2−ΔΔCT method to assess the levels of mRNAs encoding targeted genes normalized to GAPDH.