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JoVE Journal Genetics
Transcriptome-Wide Profiling of Protein-RNA Interactions by Cross-Linking and Immunoprecipitation Mediated by FLAG-Biotin Tandem Purification
 

Transcriptome-Wide Profiling of Protein-RNA Interactions by Cross-Linking and Immunoprecipitation Mediated by FLAG-Biotin Tandem Purification

Article DOI: 10.3791/60730-v • 08:46 min • May 18th, 2020
May 18th, 2020 •
1,2, 1, 1
1Tsinghua-Peking Center for Life Sciences, School of Medicine and School of Life Sciences, Tsinghua University, 2Department of Molecular Biology, University of Texas Southwestern Medical Center

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Here we present a modified CLIP-seq protocol called FbioCLIP-seq with FLAG-biotin tandem purification to determine the RNA targets of RNA-binding proteins (RBPs) in mammalian cells.

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Transcriptome-Wide Profiling Protein-RNA Interactions Cross-Linking And Immunoprecipitation FLAG-Biotin Tandem Purification Binding Proteins CLIP-seq Method Special And Temporal Arrangement Of RNAs And RBPs Mammalian Cells Direct Protein-bound RNAs SDS-PAGE Membrane Transfer Procedures Isotope Free CLIP-seq Protein Specific Antibody Mouse Embryonic STEM Cells Trypsin EDTA MESC Medium Centrifuge Tube Cross-linking UV Light Radiation Cell Lysis
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