Products/Services Used | Details | Operation |
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PCR Cloning and Subcloning> | Full-length nucleocapsid construct was cloned into a pESPRIT vector between the Aat II and Not I cleavage sites with His6-tag and TEV protease cleavage sites at the N terminus (GenScript Biotech, The Netherlands). | Get A Quote |
Many biomolecular condensates are enriched in and depend on RNAs and RNA binding proteins (RBPs). So far, only a few studies have addressed the characterization of the intermolecular interactions responsible for liquid-liquid phase separation (LLPS) and the impact of condensation on RBPs and RNAs. Here, we present an approach to study protein-RNA interactions inside biomolecular condensates by applying cross-linking of isotope labeled RNA and tandem mass spectrometry to phase-separating systems (LLPS-CLIR-MS). LLPS-CLIR-MS enables the characterization of intermolecular interactions present within biomolecular condensates at residue-specific resolution and allows a comparison with the same complexes in the dispe... More