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A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
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Genetics
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JoVE Journal Genetics
A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools

A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools

DOI:
10.3791/59901-v

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00:09 min

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August 21, 2019

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, ,
1Division of Medical Genetics,University of Washington School of Medicine

Chapters

  • 00:04Title
  • 00:513’ Adaptor Ligation
  • 04:465’ Linker Ligation
  • 05:44Reverse Transcription (RT)
  • 06:33PCR Amplification and Agarose Gel Purification
  • 08:10Results: Representative MicroRNA Analyses
  • 08:49Conclusion

Summary

Automatic Translation

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Here, we describe a step-by-step strategy for isolating small RNAs, enriching for microRNAs, and preparing samples for high-throughput sequencing. We then describe how to process sequence reads and align them to microRNAs, using open source tools.

Tags

Keywords: MicroRNARNA SequencingBioinformaticsGel PurificationSmall RNARNA IsolationGene TherapyDisease Mechanism

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