Protocol for Disome-seq to identify transcriptome-wide ribosome collisions in yeast cells

When translation elongation is hindered, stalled ribosomes can collide with trailing ribosomes and form ribosome collision complexes also known as disomes. Disomes are sensed in the cell to trigger signaling events, and therefore it is important to determine their frequency and distribution across transcripts. Here, we provide the protocol for Disome-seq in Saccharomyces cerevisiae (yeast) cells to enable transcriptome-wide detection of disomes. We describe the steps for yeast growth and isolation of disomes as well as sequencing library preparation. For complete details on the use and execution of the protocol, please refer to Meydan et al.¹ and Meydan et al.².