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Internal Reference Gene Selection for Quantitative Real-Time RT-PCR Normalization in Potato Tissues

Gang Li1, Yao Zhou2, Yaqi Zhao2, Yaxue Liu2, Yuwei Ke2, Xiaoqing Jin1, Haoli Ma1,2,*

1 Department of Biological Repositories, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China
2 College of Agronomy, Northwest A&F University, Yangling, Shaanxi, 712100, China

* Corresponding Author: Haoli Ma. Email: email

Phyton-International Journal of Experimental Botany 2020, 89(2), 329-344. https://doi.org/10.32604/phyton.2020.08874

Abstract

Quantitative real-time PCR (qRT-PCR) is widely used for investigating gene expression patterns and has many advantages, including its high sensitivity, fidelity, and specificity. Selecting a satisfactory internal reference gene is crucial for obtaining precise gene expression results in qRT-PCR analyses. In this study, the transcriptomic data of 2 potato varieties were screened for housekeeping genes with stable expression patterns. A total of 77 putative genes were selected, which were highly and stably expressed. Then, qRT-PCR analyses were performed to examine the expression levels of these 77 candidate reference genes in various potato tissues, including leaves, flowers, stolons, and tubers. Gene expression was represented by analyzing the Ct values at given threshold. Through geNorm and NormFinder program analyses, 10 candidate genes with the most stable expression patterns were obtained, including RPL19, RPS15, RPS9, EF1α, TrxP1, RPS8, NTF, CAM, AACM, and RPS28. Moreover, through the comprehensive analyses of 4 statistical algorithms (i.e., geNorm, NormFinder, BestKeeper, and RefFinder), results indicated that the most appropriate internal reference genes were RPL19 and EF1α. The obtained stable reference genes will contribute to future qRT-PCR analyses on potato tissue-related gene expression.

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APA Style
Li, G., Zhou, Y., Zhao, Y., Liu, Y., Ke, Y. et al. (2020). Internal reference gene selection for quantitative real-time RT-PCR normalization in potato tissues. Phyton-International Journal of Experimental Botany, 89(2), 329-344. https://doi.org/10.32604/phyton.2020.08874
Vancouver Style
Li G, Zhou Y, Zhao Y, Liu Y, Ke Y, Jin X, et al. Internal reference gene selection for quantitative real-time RT-PCR normalization in potato tissues. Phyton-Int J Exp Bot. 2020;89(2):329-344 https://doi.org/10.32604/phyton.2020.08874
IEEE Style
G. Li et al., "Internal Reference Gene Selection for Quantitative Real-Time RT-PCR Normalization in Potato Tissues," Phyton-Int. J. Exp. Bot., vol. 89, no. 2, pp. 329-344. 2020. https://doi.org/10.32604/phyton.2020.08874

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cc This work is licensed under a Creative Commons Attribution 4.0 International License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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