library_construction_protocol: The fragmented RNA served as a template for first-strand cDNA synthesis using random hexamer primers. Following the removal of the RNA template, a replacement strand was synthesized, creating double-stranded cDNA with dUTP in place of dTTP. The second strand was quenched during amplification by the incorporation of dUTP. To avoid self-ligation during adapter ligation, a single 'A' nucleotide was added to the 3' ends of fragments. Multiple indexing adapters were ligated to the prepared double-stranded cDNA, subsequently selecting cDNA fragments ranging from 300 bp through size selection. Phusion DNA polymerase enabled PCR amplification, generating final libraries that were quantified using TBS380 and subjected to paired-end sequencing on an Illumina novaseq6000 sequencer with 2x150 bp read length.