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Use of Alu Element Containing Minigenes to Analyze Circular RNAs
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Biochemistry
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JoVE Journal Biochemistry
Use of Alu Element Containing Minigenes to Analyze Circular RNAs

Use of Alu Element Containing Minigenes to Analyze Circular RNAs

DOI:
10.3791/59760-v

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13:10 min

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March 10, 2020

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, , ,
1Institute for Molecular and Cellular Biochemistry,University of Kentucky

Chapters

  • 00:05Introduction
  • 01:09Design of the Constructs
  • 02:14Select the Sequence to be Cloned in an Expression Vector
  • 03:35Design Primers for Cloning
  • 04:59Cloning of Constructs
  • 07:05Transfection of the Reporter Genes into Eukaryotic Cells
  • 10:21Results: Analysis of Circular RNAs
  • 11:09Conclusion

Summary

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We clone and analyze reporter genes generating circular RNAs. These reporter genes are larger than constructs to analyze linear splicing and contain Alu elements. To investigate the circular RNAs, the constructs are transfected into cells and resulting RNA is analyzed using RT-PCR after removal of linear RNA.

Tags

Circular RNAAlternative SplicingGenomic Expression SystemAlu ElementPCR OptimizationUCSC Genome BrowserRepetitive ElementsPrimer DesignExon-intron BordersReverse Complement

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