Sybr green hiscript 2 q rt supermix for qpcr kit

Total RNA was extracted from the collected samples with Total RNA Kit I (OMEGA, R6834-02). The complementary DNA was synthesized using ClonExpress Ultra One Step Cloning Kit (Vazyme, C115-02) according to the manufacturer’s instructions. Quantitative PCR reaction mixture was prepared using SYBR Green HiScript II Q RT SuperMix for qPCR Kit (Vazyme, R223-01). The qPCR reaction was performed on ABI ViiA7 Real-Time thermal cycler (Thermo Fisher, ABI). Primers for qPCR were listed in Supplementary Table 1. The relative expression of gene of interest was calculated using the 2^-ΔΔCT method [47 (link)]. qPCR analysis was performed with three technical replicates. The data represent the means ± standard errors (n = 3). P value < 0.05 was considered as significant difference.

Total RNA was isolated from cells using Total RNA Kit I (OMEGA, R6834-02) according to the manufacturer’s instructions. cDNA was synthesized using ClonExpress Ultra One Step Cloning Kit (Vazyme, C115-02) followed by qRT-PCR using SYBR Green HiScript II Q RT SuperMix for qPCR Kit (Vazyme, R223-01). Real-time quantitative RT-PCR analysis was performed using ABI ViiA7 Real-Time thermal cycler (Thermo Fisher, ABI). The primers used in the study were synthesized in Sangon Biotech (Guangzhou, China) and showed in Supplementary Table 2. The mRNA expression levels were normalized to β-actin, and fold induction was calculated by the ΔΔCT method. RT-qPCR was performed in triplicate.