Abstract
The quantitative real-time polymerase chain reaction is used to simultaneously amplify and quantify a targeted DNA molecule. It can be used to determine exact copy number of a molecule within a sample and/or to compare the quantity of a molecule between samples. When combined with reverse transcription, it is a powerful tool for the analysis of gene expression, and it is widely used for this purpose in plant species. Here we provide an introduction to fundamental concepts relevant for the analysis of gene expression in plants using this technique and a protocol for quantification of the relative expression of a sucrose phosphate synthase gene along the maturation gradient of a sugarcane leaf.
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Acknowledgements
We wish to acknowledge the Australian Research Council and the Grains Research and Development Corporation for Funding and Dr Rosanne Casu (CSIRO Plant Industry, Australia) for critical reading.
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Fitzgerald, T.L., McQualter, R.B. (2014). The Quantitative Real-Time Polymerase Chain Reaction for the Analysis of Plant Gene Expression. In: Henry, R., Furtado, A. (eds) Cereal Genomics. Methods in Molecular Biology, vol 1099. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-715-0_9
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DOI: https://doi.org/10.1007/978-1-62703-715-0_9
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