hek293t_jev_SM25-rep1
HEK 293T cells, JEV-infected HEK 293T cells were collected using TRIzol, and total RNA was extracted from the cells using an RNA Easy Fast Tissue/Cell Kit according to the manufacturer's instructions. The RNA was stored at -80C for further experiments. DNA primers were synthesized by Sangon Biotech (Shanghai, China). The DNA fragments were amplified via PCR using forward primers, reverse primers and 2Fast Pfu Master Mix (Novoprotein, China), with water added to a total volume of 50 L. The PCR products were purified with a SanPrep Column PCR Product Purification Kit (Sangon Biotech, China) according to the manufacturer's instructions. Then, the DNA fragments were reverse transcribed into RNA fragments using a T7 High Efficiency Transcription Kit (TransGen Biotech, China), and the reverse transcription products were purified with an EasyPure RNA Purification Kit (TransGen Biotech, China) according to the manufacturer's instructions. Then, the purified reverse transcription products were diluted to 1012 copies/L and stored at -80C for further experiments.