Hifair 3 1st strand cdna synthesis supermix for qrt pcr

As described before [16 (link)], Trizol reagent (GLPBIO, Montclair, CA, USA) was used to isolate the total RNA of liver samples, which was reverse-transcribed into cDNA by the Hifair^® III 1st Strand cDNA Synthesis SuperMix for qRT-PCR (gDNA digester plus) according to the instruction (Yeasen, Shanghai, China). The cDNA obtained in the experiment was perfromed by HRbio™ qPCR SYBR Green Master Mix (Fujian Herui Biotechnology, Fuzhou, China). The primer sequences used were listed in Table 1.

RNA was extracted with the RNAiso Plus reagent (Takara Bio, Shiga, Japan, catalog no. 9109) and reverse-transcribed with Hifair III 1st Strand cDNA Synthesis SuperMix for qRT-PCR (Yeasen Biotechnology, Shanghai, China, catalog no. 11141ES60). The reverse transcription products of different samples were amplified by a LightCycler System (Roche, Mannheim, Germany) using the SYBR Green Master Mix (Yeasen Biotechnology, Shanghai, China, catalog no. 11201ES08) according to the manufacturer’s instruction, and data were normalized by the level of GAPDH in each sample. The 2^-ΔΔCt method was used to calculate relative expression changes. With the help of the dissociation curve analysis, specific primer pairs of every gene were designed and selected. The sequences of the primers for qRT-PCR are shown in Supplemental Table S1. All experiments were repeated at least three times.