Genome-wide sequencing of RNA (RNA-seq) has become an inexpensive tool to gain key insights into cellular and disease mechanisms. Sample preparation and sequencing are streamlined and allow the acquisition of hundreds of gene expression profiles in a few days; however, in particular, data processing, curation, and analysis involve numerous steps that can be overwhelming to non-experts. Here, the sample preparation, sequencing, and data processing workflow for RNA-seq expression analysis in yeast is described. While this protocol covers only a small portion of the RNA-seq landscape, the principal workflow common to such experiments is described, allowing the reader to adapt the protocol where necessary. Graphic abstract: Basic workflow of RNA-seq expression analysis.
Keywords: Next-generation sequencing; Relative expression levels; Sequence analysis; Systems biology; Whole genome; Yeast; mRNA.
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