design: Total RNA is extracted from tissues and then examined using devices like the Nanodrop 2000. Samples are selected according to specific requirements. Subsequently, mRNA is enriched via Oligo dT, followed by fragmentation, reverse - transcription to synthesize cDNA, and adapter ligation. After that, fragments are screened and the library is enriched. Finally, the library is quantified using a Qubit 4.4, amplified on a cBot, and sequenced on the NovaSeq X Plus platform.