RNA sequencing (RNA-Seq) is a widely used experimental approach in molecular biology. RNA-Seq allows us to measure the level of RNA in cells and tissues, and therefore make inferences about what genes are active, under what circumstances, and when. Since every cell’s copy of its genetic blueprint is the same, it is the timing and amount of gene expression—and subsequently the production of proteins—that differentiate cells by developmental time point and developmental fate. Analysis of an RNA-Seq experiment requires sophisticated bioinformatics tools to evaluate data and generate a statistical analysis of gene expression. Due to the size and complexity of RNA-Seq datasets, these experiments are generally done on cloud computers or high-performance supercomputers. Join Jason Williams for this three-part course to learn the basics of RNA-Seq and how to use the online platform DNA Subway to analyze an example data set.