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小RNA分子的高效结扎的微小RNA定量高通量测序
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JoVE Journal Biology
Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing
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小RNA分子的高效结扎的微小RNA定量高通量测序

DOI:
10.3791/52095-v

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14:15 min

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November 18, 2014

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1Molecular, Cellular, and Developmental Biology,University of Colorado, Boulder, 2Linda Crnic Institute for Down Syndrome,University of Colorado, Denver

Chapters

  • 00:05Title
  • 01:45Adenylation of 3′ Linker
  • 05:14Linker Ligation to 3′ End of miRNA
  • 10:29Linker Ligation to 5′ End of miRNA-3′ Linker Hybrid
  • 11:46Results: Ligation of Small RNA Molecules by the Method is Highly Efficient
  • 13:50Conclusion

Summary

Automatic Translation

Automatic Translation

MicroRNAs (miRNAs) are a widely conserved class of regulatory molecules. Here we describe a miRNA cloning method that relies upon two potent ligation steps followed by high-throughput sequencing. Our method permits accurate genome-wide quantitation of miRNAs.

Tags

Keywords: MiRNAMiR-SeqHigh-throughput SequencingSmall RNA LigationLibrary ConstructionQuantitationBias ReductionTranscriptome-wide Profiling

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