Hifair 1st strand cdna synthesis supermix for qpcr gdna digester plus kit

Total RNA was isolated from the muscle sample (100 mg) using TRIzol TM Reagent (Thermo Fisher Scientific, Waltham, MA, USA), and the concentration and purity were measured according to the ratio of A260/A280 using an Epoch Microplate Spectrophotometer (BioTek Instruments, Inc., Winooski, VT, USA). Two µL of total RNA were reversely transcribed into cDNA using the Hifair^® 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) kit (Yeasen, Shanghai, China). The Hieff^® qPCR SYBR Green Master Mix (Low Rox Plus) kit (Yeasen, Shanghai, China) and Real-Time PCR System (Thermo Fisher Scientific, MA, USA) were used to quantify the mRNA levels of genes. The primers for Nrf2, kelch-like epichlorohydrin-associated protein 1 (Keap1), HO-1, NAD(P)H/quinone oxidoreductase 1 (NQO1), CAT, SOD1, GST, GSH-Px, and β-actin were synthesized by SangonBiotechCo., Ltd. (Shanghai, China) and are presented in Table S2, as in our previous study [16 (link)]. The reaction condition and parameter setting were according to the settings in our previous research [19 (link)]. The target genes’ relative mRNA expression was determined using the 2^−∆∆Ct method against β-actin as the housekeeping gene.