| Chapter title |
Global RT-PCR and RT-qPCR Analysis of the mRNA Expression of the Human PTPome
|
|---|---|
| Chapter number | 2 |
| Book title |
Protein Tyrosine Phosphatases
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3746-2_2 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3744-8, 978-1-4939-3746-2
|
| Authors |
Caroline E. Nunes-Xavier, Rafael Pulido, Nunes-Xavier, Caroline E., Pulido, Rafael |
| 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. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 4 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Bachelor | 2 | 50% |
| Student > Ph. D. Student | 1 | 25% |
| Researcher | 1 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 3 | 75% |
| Unknown | 1 | 25% |