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Global RT-PCR and RT-qPCR Analysis of the mRNA Expression of the Human PTPome

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Protein Tyrosine Phosphatases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1447))

Abstract

Comprehensive comparative gene expression analysis of the tyrosine phosphatase superfamily members (PTPome) under cell- or tissue-specific growth conditions may help to define their individual and specific role in physiology and disease. Semi-quantitative and quantitative PCR are commonly used methods to analyze and measure gene expression. Here, we describe technical aspects of PTPome mRNA expression analysis by semi-quantitative RT-PCR and quantitative RT-PCR (RT-qPCR). We provide a protocol for each method consisting in reverse transcription followed by PCR using a global platform of specific PTP primers. The chapter includes aspects from primer validation to the setup of the PTPome RT-qPCR platform. Examples are given of PTP-profiling gene expression analysis using a human breast cancer cell line upon long-term or short-term treatment with cell signaling-activation agents.

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Acknowledgements

This work was supported in part by grants SAF2009-10226 from Ministerio Ciencia e Innovación (Spain and Fondo Europeo de Desarrollo Regional), SAF2013-48812-R from Ministerio de Economía y Competitividad (Spain), and EU Research Training Network (MRTN-CT-2006-035830).

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Authors and Affiliations

  1. Centro de Investigación Príncipe Felipe, 46013, Valencia, Spain

    Caroline E. Nunes-Xavier

  2. Department of Tumor Biology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, 4950, Nydalen, 0424, Oslo, Norway

    Caroline E. Nunes-Xavier

  3. Biocruces Health Research Institute, Pza Cruces s/n, 48903, Barakaldo, Spain

    Caroline E. Nunes-Xavier & Rafael Pulido

  4. IKERBASQUE, Basque Foundation for Science, 48013, Bilbao, Spain

    Rafael Pulido

Authors
  1. Caroline E. Nunes-Xavier
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  2. Rafael Pulido
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Corresponding authors

Correspondence to Caroline E. Nunes-Xavier or Rafael Pulido .

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Editors and Affiliations

  1. BioCruces Health Research Institute, Barakaldo, Bizkaia, Spain

    Rafael Pulido

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Nunes-Xavier, C.E., Pulido, R. (2016). Global RT-PCR and RT-qPCR Analysis of the mRNA Expression of the Human PTPome. In: Pulido, R. (eds) Protein Tyrosine Phosphatases. Methods in Molecular Biology, vol 1447. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3746-2_2

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  • DOI: https://doi.org/10.1007/978-1-4939-3746-2_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3744-8

  • Online ISBN: 978-1-4939-3746-2

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