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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
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Genetics
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JoVE Journal Genetics
An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells

An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells

DOI:
10.3791/59628-v

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09:45 min

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May 21, 2019

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, , ,
1Department of Biomedical Science and Research Institute for Bioscience & Biotechnology,Hallym University

Chapters

  • 00:04Title
  • 01:06MicroRNA (miRNA) Mimic Transfection
  • 02:03Sulforhodamine B (SRB) Assay
  • 03:20Generation of a Dose-response Curve
  • 04:35Verification of the Direct Target Gene of a miRNA of Interest
  • 06:37Luciferase Assay
  • 07:52Results: miRNA-107 Negatively Regulates Proliferation of PANC-1 Cells
  • 08:53Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol uses a probe-based real-time polymerase chain reaction (PCR), a sulforhodamine B (SRB) assay, 3’ untranslated regions (3’ UTR) cloning, and a luciferase assay to verify the target genes of a miRNA of interest and to understand the functions of miRNAs.

Tags

Keywords: MicroRNAMicroRNA TargetIn Vitro ProtocolTumor CellsCell Viability AssayTransfectionMicroRNA MimicMicroRNA-107

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