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

  • Marie P. Khoury
  • Virginie Marcel
  • Kenneth Fernandes
  • Alexandra Diot
  • David P. Lane
  • Jean-Christophe Bourdon
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

p53 isoforms Cancer Detection Quantification mRNA Splicing Nested PCR Quantitative real-time RT-PCR (TaqMan® chemistry) p53 tumor suppressor protein 

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Marie P. Khoury
    • 1
  • Virginie Marcel
    • 1
  • Kenneth Fernandes
    • 1
  • Alexandra Diot
    • 1
  • David P. Lane
    • 1
  • Jean-Christophe Bourdon
    • 2
  1. 1.CR-UK Cell Transformation Research Group, Centre for Oncology and Molecular Medicine, Ninewells HospitalUniversity of DundeeDundeeUK
  2. 2.Department of Surgery and Molecular Oncology, Ninewells HospitalUniversity of DundeeDundeeUK

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