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Confirmation of Transcriptional Read-Through Events by RT-PCR

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Chimeric RNA

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2079))

Abstract

Chimeric RNAs can be formed by trans-splicing from different transcripts or cis-splicing of adjacent genes (cis-SAGe). Cis-SAGe results from read-through transcription of two neighbor genes. To investigate the mechanisms underlying intergenic splicing of adjacent genes, it is important to develop an assay to detect transcriptional read-through. Here, we describe a general RT-PCR based method to confirm the process for cis-SAGe candidates. In this method, we use PCR to amplify cDNA that is reverse transcribed from the read-through precursor mRNA. The result provides a foundation for further downstream mechanistic studies.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Genetics, School of Medicine, University of Virginia, Charlottesville, VA, USA

    **nrui Shi

  2. Department of Pathology, School of Medicine, University of Virginia, Charlottesville, VA, USA

    Fujun Qin & Hui Li

Authors
  1. **nrui Shi
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  2. Fujun Qin
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  3. Hui Li
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Corresponding author

Correspondence to Hui Li .

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Editors and Affiliations

  1. Department of Paathology, School of Medicine, University of Virginia, Charlottesville, VA, USA, Department of Biochemistry and Molecular Genetics, School of Medicine, University of Virginia, Charlottesville, VA, USA

    Hui Li

  2. Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA, USA

    Justin Elfman

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Shi, X., Qin, F., Li, H. (2020). Confirmation of Transcriptional Read-Through Events by RT-PCR. In: Li, H., Elfman, J. (eds) Chimeric RNA. Methods in Molecular Biology, vol 2079. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9904-0_14

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  • DOI: https://doi.org/10.1007/978-1-4939-9904-0_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9903-3

  • Online ISBN: 978-1-4939-9904-0

  • eBook Packages: Springer Protocols

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