Purification of Ribosome-Nascent-Chain Complex for Ribosome Profiling and Selective Ribosome Profiling.

Hagit Bar-Yosef, Johannes Venezian, Kevin Klann, Ayala Shiber

Methods in molecular biology (Clifton, N.J.) 2022

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Selective Ribosome Profiling (SeRP) is an emerging methodology, developed to capture cotranslational interactions in vivo. To date, SeRP is the only method that can directly capture, in near-codon resolution, ribosomes in action. Thus, SeRP allows us to study the mechanisms of protein synthesis and the network of protein-protein interactions that are formed already during synthesis. Here we report, in detail, the protocol for purification of ribosome- and Nascent-Chain associated factors, followed by isolation of ribosome-protected mRNA footprints, cDNA library generation and subsequent data analysis. © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Hagit Bar-Yosef, Johannes Venezian, Kevin Klann, Ayala Shiber. Purification of Ribosome-Nascent-Chain Complex for Ribosome Profiling and Selective Ribosome Profiling. Methods in molecular biology (Clifton, N.J.). 2022;2477:179-193