title: Global RT-PCR and RT-qPCR Analysis of the mRNA Expression of the Human PTPome.
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.
authors: Centro de Investigación Príncipe Felipe, 46013, Valencia, Spain. carnun@rr-research.no.|Department of Tumor Biology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, 4950, Nydalen, 0424, Oslo, Norway. carnun@rr-research.no.|Biocruces Health Research Institute, Pza Cruces s/n, 48903, Barakaldo, Spain. carnun@rr-research.no.
authors: Caroline E
authors: CE
authors: Nunes-Xavier
authors: Biocruces Health Research Institute, Pza Cruces s/n, 48903, Barakaldo, Spain. rpulidomurillo@gmail.com.|IKERBASQUE, Basque Foundation for Science, 48013, Bilbao, Spain. rpulidomurillo@gmail.com.
authors: Rafael
authors: R
authors: Pulido
mesh_terms: D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D061986:MCF-7 Cells; D017027:Protein Tyrosine Phosphatases / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D059467:Transcriptome
chemical_list: D012333:RNA, Messenger; D017027:Protein Tyrosine Phosphatases
keywords: PTPome; Protein tyrosine phosphatase; Real-time quantitative PCR; Reverse transcription PCR