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A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA
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Biologia
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JoVE Journal Biologia
A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA

DOI:
10.3791/1622-v

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13:00 min

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December 02, 2009

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, ,
1Department of Molecular and Cellular Biology,Brown University, 2Center for Computational Molecular Biology,Brown University

Capitoli

  • 00:00Titolo
  • 00:42Introduction
  • 01:17Pool Design and Oligo Recovery
  • 03:53Co-immunoprecipitation of the Oligo Pool with an RNP of Interest
  • 07:50Labeling the Samples and Hybridizations to the Microarray
  • 10:50Mapping RNPs on Pre-mRNAs Representative Results
  • 11:56Conclusion

Summary

Traduzione automatica

Traduzione automatica

Due to the transient nature of pre-mRNA, it can be difficult to isolate and study in vivo. Here, we present a novel in vitro approach to investigate RNA-protein interactions using a synthetic oligo pool that tiles across selected regions of pre-mRNA.

Tags

Rapid High-throughput MethodMapping RibonucleoproteinsRNPsHuman Pre-mRNACo-immunoprecipitateRNA Binding ProteinsSplicing FactorIn Vitro ApproachOligonucleotide PoolMolecular SelectionTwo Color Array StrategyEnrichmentHigh-resolution MapsSequence-specific DeterminatesStructural DeterminatesRibonucleoprotein BindingUnique AdvantageSampling BiasIP TechniquesSELEX TechniquesFlexibility

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