This study enriched mRNA from total RNA using Oligo(dT) magnetic beads to capture polyadenylated transcripts. RNA fragmentation was achieved through ion-based shearing, producing approximately 300 base pairs (bp) fragments. The choice of 300 bp as the fragment length was based on the following considerations: shorter fragments would result in an overrepresentation of adapter sequences, thus reducing the proportion of valid data. In comparison, longer fragments could hinder efficient cluster formation during sequencing. cDNA synthesis was carried out using RNA as a template, with a six-base random primer and reverse transcriptase for the first-strand synthesis. The second strand of cDNA was then synthesized using the first strand as a template. Following cDNA synthesis, PCR constructed and amplified the library to enrich for desired fragment sizes. Library selection was performed based on fragment size, with a target size of 450 bp.