library_construction_protocol: Total RNA extraction by trizol method (1) Eukaryotic mRNA was enriched with magnetic beads with Oligo (dT); (2) Add Fragmentation Buffer to interrupt the mRNA randomly; (3) Synthesis of the first cDNA strand and double strand using mRNA as template and cDNA purification; (4) The purified double-stranded cDNA was then subjected to end repair, addition of A-tail and ligation of sequencing junctions, followed by fragment size selection with AMPure XP beads; (5) Finally, the cDNA library was enriched by PCR.