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

Autor: Hagit, Bar-Yosef, Johannes, Venezian, Kevin, Klann, Ayala, Shiber
Rok vydání: 2022
Předmět:
Zdroj: Methods in molecular biology (Clifton, N.J.). 2477
ISSN: 1940-6029
Popis: 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.
Databáze: OpenAIRE