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

  • Caroline E. Nunes-XavierEmail author
  • Rafael PulidoEmail author
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Protein tyrosine phosphatase Reverse transcription PCR Real-time quantitative PCR PTPome 

Notes

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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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Centro de Investigación Príncipe FelipeValenciaSpain
  2. 2.Department of Tumor Biology, Institute for Cancer ResearchOslo University Hospital RadiumhospitaletOsloNorway
  3. 3.Biocruces Health Research InstituteBarakaldoSpain
  4. 4.IKERBASQUEBasque Foundation for ScienceBilbaoSpain

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