Identification and Validation of Reliable Reference Genes for Gene Expression Studies in .

Genes (Basel)

Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, National Engineering Research Center of Tree breeding and Ecological Restoration, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China.

Published: April 2022

RT-qPCR is considered a rapid and reliable technique for analyzing gene expression. This technique is commonly used to analyze the expression of various genes at diverse transcriptional levels in different samples. However, few studies have characterized ornamental species for reliable reference genes. In this study, eight reference genes were evaluated as controls in RT-qPCR with SYBR green to quantify gene expression in different samples. All selected reference genes showed a broad range of C values in all samples, which was supportive of their variable expression. Our results showed significant variation in the stable expression of genes. Sample data, analyzed using geNorm, NormFinder, and BestKeeper, showed that phospholipase () and β-actin () were the most suitable and statistically reliable reference genes, whereas ribosomal protein L13 () and elongation factor 1-α () were less stable and unsuitable for use as internal controls. To compare gene expression levels, two or more reference genes should be used for data normalization. Thus, the stability and expression of both and were believed to provide better normalization and quantification of the transcript levels for gene expression studies in .

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9141280PMC
http://dx.doi.org/10.3390/genes13050714DOI Listing

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