Abstract
With a widely established use of quantitative real-time PCR (qRT-PCR) for gene expression analysis, reliable and stable expression of reference genes is often discussed. Suitable reference genes should show less variation of expression across the target samples and allow for error minimization by normalization of qRT-PCR data. Therefore, selection of reliable reference genes is essential for accurate results and to support the conclusions drawn on expression levels of genes under study. In this chapter, we describe the workflow for selection and evaluation of reference genes in rice, including identification of candidate genes by using Genevestigator® and evaluation of expression stability using various algorithms. The ranking of the genes guides qRT-PCR performance and data analysis. This protocol used rice as an example but is not limited to rice, and could be applied to other species as well.
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Acknowledgments
This research was supported by funds from State Secretariat for Education, Research and Innovation to N.K.B. We thank Prof. Gruissem for providing access to Lab infrastructure. We thank Irene Zurkirchen for the technical support in the greenhouse.
Conflict of Interest Statement: None declared.
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Wang, M., Bhullar, N.K. (2021). Selection of Suitable Reference Genes for qRT-PCR Gene Expression Studies in Rice. In: Bandyopadhyay, A., Thilmony, R. (eds) Rice Genome Engineering and Gene Editing. Methods in Molecular Biology, vol 2238. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1068-8_20
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DOI: https://doi.org/10.1007/978-1-0716-1068-8_20
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