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Quantitative Reverse Transcription-qPCR-Based Gene Expression Analysis in Plants

  • Heithem Ben Abdallah
  • Petra BauerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1363)

Abstract

The investigation of gene expression is an initial and essential step to understand the function of a gene in a physiological context. Reverse transcription-quantitative real-time PCR (RT-qPCR) assays are reproducible, quantitative, and fast. They can be adapted to study model and non-model plant species without the need to have whole genome or transcriptome sequence data available. Here, we provide a protocol for a reliable RT-qPCR assay, which can be easily adapted to any plant species of interest. We describe the design of the qPCR strategy and primer design, considerations for plant material generation, RNA preparation and cDNA synthesis, qPCR setup and run, and qPCR data analysis, interpretation, and final presentation.

Key words

Gene expression Primer design Reverse transcription cDNA qPCR Reference gene Cq value 

Notes

Acknowledgements

H.B.A. was supported through internship and STIBET fellowships from the DAAD and the Tunisian government.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Institute of BotanyHeinrich-Heine UniversityDüsseldorfGermany
  2. 2.Center of Excellence in Plant Sciences (CEPLAS)DüsseldorfGermany

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