RNA-sequencing (RNA-seq) is a ubiquitous tool in modern molecular biology. It greatly helps us to understand genomic functions and transcriptional changes occurring in disease states, in response to therapeutics and under different environmental conditions. The goal for RNA-seq analysis is to identify differentially expressed genes (DEG) and pathways that are related to a certain condition. In this seminar, we will cover the basic concepts in RNA-seq, QC, filtering, normalization, DEG analysis, and enrichment analysis using R packages from Bioconductor. We will go through the workflow using a dataset from a published manuscript.