Transscript first stand cdna synthesis kit

Total RNA was isolated from the cultured cells using an Eastep Total RNA Extraction kit (Promega) according to the manufacturer’s protocol. Then, total RNA was reverse transcribed using a TransScript First-Stand cDNA Synthesis kit (TransGen Biotech). RT-qPCR was subsequently performed with TransStart Green qPCR SuperMix (TransGen Biotech), and products were detected with a DA7600 Real-time Nucleic Acid Amplification Fluorescence Detection System (Bio-Rad, Hercules, CA, USA). We quantified the transcripts of the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal mRNA quantity control. All primers for cDNA amplification of various target genes were designed and optimized using Oligo 7.0 software (Molecular Biology Insights, West Cascade, USA) and synthesised by Sangon Biotech (Shanghai, China). The relative expression levels of the target genes were calculated by normalizing the cycle threshold (Ct) values of the target gene to the Ct values of GAPDH (ΔCt) and determined as 2^-ΔCt. The RT-qPCR products were subjected to electrophoresis.