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Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells
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Genetics
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JoVE Journal Genetics
Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells

Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells

DOI:
10.3791/58655-v

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10:55 min

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March 08, 2019

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, , , , ,
1The Interdisciplinary Nanoscience Centre,Aarhus University, 2Department of Molecular Biology and Genetics,Aarhus University

Chapters

  • 00:04Title
  • 00:33Cell Culture, and the Collection of EVs and RNA-associated Biomolecules
  • 02:54RNA Extraction and Quality Control
  • 04:20Library Construction and Quality Control
  • 09:04Results: Collecting exRNA from hMSC for Next Generation Sequencing
  • 10:31Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol demonstrates how to purify extracellular microRNAs from cell culture media for small RNA library construction and next generation sequencing. Various quality control checkpoints are described to allow readers to understand what to expect when working with low input samples like exRNAs.

Tags

Keywords: Extracellular MicroRNAsMesenchymal Stem CellsNext-generation SequencingExtracellular VesiclesRNA IsolationNanoparticle Tracking AnalysisUltracentrifugationQuality Control

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