RNA-Seq of DEF Cells:with viral infection
After extracting the sample total RNA and digesting the DNA using DNase, eukaryotic mRNA is enriched with magnetic beads with Oligo(dT); The mRNA was broken into short fragments by adding interrupting reagent, and the interrupted mRNA was used as a template to synthesize one-strand cDNA with six-base random primers, and then the two-stranded cDNA was synthesized into a two-strand reaction system, and the double-stranded cDNA was purified by the kit. The purified double-stranded cDNA was then repaired at the end, A-tailed and ligated with sequencing adapters, followed by fragment size selection, and finally PCR amplification. After the libraries were qualified with the Agilent 2100 Bioanalyzer, they were sequenced using an Illumina HiSeq2500 or Illumina HiSeq X Ten, resulting in 125 bp or 150 bp paired-end data. After passing the quality check, the sequencing was performed using an Illumina sequencer.