Guidelines for performing Ribosome Profling in Plants Including Structural Analysis of rRNA fragments

Here, we compile valuable insights gathered over years of generating Ribo-seq datasets from different plant species and experimental setups. We tested the effects of variable ribonuclease treatments for the generation of ribosome protected fragments (RPFs). We tested rRNA depletion strategies designed specifically for Arabidopsis and Tobacco. We also compare ligation-free to ligation-based library preparation strategies for generating Ribo-seq libraries. Overall design: RPFs were generated following different nuclease treatments, followed by the preparation of Ribo-seq libraries.