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Detecting and Quantifying p53 Isoforms at mRNA Level in Cell Lines and Tissues

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p53 Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 962))

Abstract

The TP53 gene expresses at least nine different mRNA variants (p53 isoform mRNAs), including the one encoding the canonical p53 tumor suppressor protein. We have developed scientific tools to specifically detect and quantify p53 isoform expression at mRNA level by nested RT-PCR (reverse transcription-polymerase chain reaction) and quantitative real-time RT-PCR (RT-qPCR using the TaqMan® chemistry). Here, we describe these two methods, while highlighting essential points with regard to the analysis of p53 isoform mRNA expression.

Marie P. Khoury and Virginie Marcel have contributed equally to this work.

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Acknowledgments

This work was supported by Cancer Research UK (C8/A6613). M.K., K.F., A.D., D.L., and J.C.B. are supported by Cancer Research UK and V.M. is supported by Breast Cancer Campaign.

Author information

Authors and Affiliations

  1. CR-UK Cell Transformation Research Group, Centre for Oncology and Molecular Medicine, Ninewells Hospital, University of Dundee, Dundee, UK

    Marie P. Khoury, Virginie Marcel, Kenneth Fernandes, Alexandra Diot & David P. Lane

  2. Department of Surgery and Molecular Oncology, Ninewells Hospital, University of Dundee, Dundee, UK

    Jean-Christophe Bourdon

Authors
  1. Marie P. Khoury
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  2. Virginie Marcel
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  3. Kenneth Fernandes
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  4. Alexandra Diot
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  5. David P. Lane
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  6. Jean-Christophe Bourdon
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Corresponding author

Correspondence to Jean-Christophe Bourdon .

Editor information

Editors and Affiliations

  1. Massey Cancer Center, Department of Biochemistry, Virginia Commonwealth University, College Street 401, Richmond, 23298, Virginia, USA

    Sumitra Deb

  2. Massey Cancer Center, Department of Biochemistry, Virginia Commonwealth University, College Street 401, Richmond, 23298, Virginia, USA

    Swati Palit Deb

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Khoury, M.P., Marcel, V., Fernandes, K., Diot, A., Lane, D.P., Bourdon, JC. (2013). Detecting and Quantifying p53 Isoforms at mRNA Level in Cell Lines and Tissues. In: Deb, S., Deb, S. (eds) p53 Protocols. Methods in Molecular Biology, vol 962. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-236-0_1

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  • DOI: https://doi.org/10.1007/978-1-62703-236-0_1

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-235-3

  • Online ISBN: 978-1-62703-236-0

  • eBook Packages: Springer Protocols

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