Transcript first strand cdna synthesis supermix

The LNCaP cells grown to the logarithmic phase were subcultured into culture dishes and divided into four groups: blank control with DMSO added, DHT-added, 30 nM rapamycin+1 nM DHT, and 30 nM rapamycin +10 nM DHT. After attached, the cells were starved for 12 h in a serum-free RPMI 1640 medium. After rapamycin of 30 nM was added for 2 h, 1 nM DHT and 10 nM DHT were added respectively to the cells for 12 h. The same volume of culture medium was added to the blank control group, and 10 nM DHT was added to the DHT control group.
Total RNA was extracted using Trizol. cDNA was synthesized and amplified using the Transcript First-Strand cDNA Synthesis SuperMix (TaKaRa Biotechnology, Dalian, China) in accordance with the manufacturer's protocol. The real-time quantitative reverse transcription PCR (qRT-PCR) was performed by SYBR Premix Ex TapTM II (TaKaRa Biotechnology, Dalian, China) and the assay was carried out in triplicate on a CFX96™ Real-Time system (Bio-Rad, Hercules, CA). The designs of primer sequences referred our previous work [27 (link)]. All reactions were performed in triplicate.