Titlebook: Nuclear Bodies and Noncoding RNAs; Methods and Protocol Shinichi Nakagawa,Tetsuro Hirose Book 2015 Springer Science+Business Media New York

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Studying RNA-Binding Protein Interactions with Target mRNAs in Eukaryotic Cells: Native Ribonucleoprction can be studied in numerous ways that range from in vitro to in vivo through messenger ribonucleoprotein immunoprecipitation (mRNP-IP or RIP) assays. This modified immunoprecipitation approach is an important and sensitive method to determine the regulation of gene expression by specific RBPs under different cellular stressors.

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Ultrastructural Analysis of Nuclear Bodies Using Electron Microscopynd tissue samples that can be re-probed whenever new antibodies or probes become available. Rapid and robust protocols are available for both I-EM and EM-ISH post-embedding techniques so that they can be combined on the same sections, providing ultrastructural and molecular insights into newly “emerging” nuclear bodies.

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Cross-Linking and Immunoprecipitation of Nuclear RNA-Binding Proteinsution. Here, we describe in detail a PAR-CLIP protocol to prepare cDNA libraries for high-throughput sequencing from RNA fragments that are bound to RBPs not only in the nucleus but also in the cytoplasm.

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https://doi.org/10.1007/978-1-4939-2253-6biochemical; cytological; eukaryotes; gene expression; long non-coding RNAs; molecular biological methods

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978-1-4939-5549-7Springer Science+Business Media New York 2015

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Shinichi Nakagawa,Tetsuro HiroseIncludes cutting-edge methods and protocols.Provides step-by-step detail essential for reproducible results.Contains key notes and implementation advice from the experts.Includes supplementary materia

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