RNA sequencing (RNA-Seq) is a powerful and widely used technique in genomics and proteomics for analyzing the transcriptome, which represents all RNA molecules, including mRNA, rRNA, tRNA, and non-coding RNAs, present in a cell at a given time. The principle of RNA-Seq is based on converting RNA into complementary DNA (cDNA), fragmenting it, and then sequencing these fragments using high-throughput sequencing technologies. This allows researchers to determine the sequence of RNA molecules, quantify gene expression levels, identify novel transcripts, alternative splicing events, and post-transcriptional modifications. RNA-Seq provides a high-resolution and comprehensive view of the transcriptome compared to traditional methods like microarrays, which are limited to detecting known sequences. The methodology of RNA sequencing involves several key steps. First, total RNA is isolated from the sample of interest, followed by the enrichment of mRNA using poly-A selection or depletion of rRNA to focus on coding RNA. The RNA is then fragmented into smaller pieces to facilitate sequencing. Reverse transcription is performed to convert the RNA fragments into cDNA. These cDNA fragments are subsequently ligated with sequencing adapters and amplified