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Titlebook: Regulatory Non-Coding RNAs; Methods and Protocol Gordon G. Carmichael Book 2015 Springer Science+Business Media New York 2015 Fluorescence

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A Biochemical Approach to Identify Direct MicroRNA Targets,y the tumor suppressor miRNA miR-34a. This method involves transfection of cells with biotinylated miRNA mimics, streptavidin pulldown, RNA isolation, and qRT-PCR. The protocol in this chapter describes these steps and the issues that should be considered while designing such pulldown experiments.
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Identification and Validation of miRNA Target Sites Within Nontraditional miRNA Targets,derstood to repress gene expression posttranscriptionally through recognition of 3′-untranslated regions (3′-UTRs) of mRNA transcripts, they have the potential to target additional classes of RNAs. Understanding the expanding pool of potential miRNA targets has been hindered by the lack of tools for
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,Affinity Purification of Long Noncoding RNA–Protein Complexes from Formaldehyde Cross-Linked Mammalsses. The purification and analysis of lncRNA–protein (lncRNP) complexes is critical toward understanding the normal physiological function of these molecules. Here, we describe the purification of lncRNP complexes from human cells using a FLAG-tagged MS2-phage coat protein (MS2 CP) that binds in se
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