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Selection and Validation of Appropriate Reference Genes for Quantitative RT-PCR Analysis in Rubia yunnanensis Diels Based on Transcriptome Data
Joint Authors
Yi, Shanyong
Lin, Qianwen
Zhang, Xuejia
Wang, Jing
Miao, Yuanyuan
Tan, Ninghua
Source
Issue
Vol. 2020, Issue 2020 (31 Dec. 2020), pp.1-19, 19 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2020-01-09
Country of Publication
Egypt
No. of Pages
19
Main Subjects
Abstract EN
Real-time quantitative polymerase chain reaction (RT-qPCR) has been widely applied in gene expression and transcription abundance analysis because of its high sensitivity, good repeatability, and strong specificity.
Selection of relatively stable reference genes is a precondition in order to obtain the reliable analysis results.
However, little is known about evaluation of a set of reference genes through scientific experiments in Rubia plants.
Here, 15 candidate reference genes were selected from R.
yunnanensis transcriptome database and analyzed under abiotic stresses, hormone treatments, and different tissues.
Among these 15 candidate reference genes, heterogeneous nuclear ribonucleoprotein (hnRNP), TATA binding protein (TBP), ribosomal protein L5 (RPL5), malate dehydrogenase (MDH), and elongation factor 1-alpha (EF-1α) were indicated as the five most stable reference genes by four statistical programs (geNorm, NormFinder, BestKeeper, and RefFinder).
Ultimately, the validity of reference genes was confirmed by normalizing the expression of o-succinylbenzoate-CoA ligase (OSBL) and isochorismate synthase (ICS) involved in the anthraquinone biosynthesis pathway in different tissues and hormone treatments.
Meanwhile, four other putative genes involved in the anthraquinone biosynthesis pathway were also normalized with the selected reference genes, which showed similar expression levels with those given by transcriptome data.
This work is the first research that aims at a systematic validation on the stability of reference genes selected from R.
yunnanensis transcriptome data and will be conducive to analyze gene expression in R.
yunnanensis or other Rubia species.
American Psychological Association (APA)
Yi, Shanyong& Lin, Qianwen& Zhang, Xuejia& Wang, Jing& Miao, Yuanyuan& Tan, Ninghua. 2020. Selection and Validation of Appropriate Reference Genes for Quantitative RT-PCR Analysis in Rubia yunnanensis Diels Based on Transcriptome Data. BioMed Research International،Vol. 2020, no. 2020, pp.1-19.
https://search.emarefa.net/detail/BIM-1134968
Modern Language Association (MLA)
Yi, Shanyong…[et al.]. Selection and Validation of Appropriate Reference Genes for Quantitative RT-PCR Analysis in Rubia yunnanensis Diels Based on Transcriptome Data. BioMed Research International No. 2020 (2020), pp.1-19.
https://search.emarefa.net/detail/BIM-1134968
American Medical Association (AMA)
Yi, Shanyong& Lin, Qianwen& Zhang, Xuejia& Wang, Jing& Miao, Yuanyuan& Tan, Ninghua. Selection and Validation of Appropriate Reference Genes for Quantitative RT-PCR Analysis in Rubia yunnanensis Diels Based on Transcriptome Data. BioMed Research International. 2020. Vol. 2020, no. 2020, pp.1-19.
https://search.emarefa.net/detail/BIM-1134968
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1134968