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Evaluation of reference genes for quantitative real-time PCR analysis of gene expression during early development processes of the tongue sole (Cynoglossus semilaevis)

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Abstract

Differential expression of genes is crucial to growth and development of fish. To select the appropriate genes for gene normalization during Cynoglossus semilaevis early developmental process, eight candidate reference genes (ACTB, B2M, EF1A, GADPH, RPL7, TUBA, UBCE and 18S) were tested for their adequacy by using quantitative real-time PCR. The results showed that the expression of all the examined genes exhibited tissue dependent variations in the mature C. semilaevis. EF1A was listed as the most stable reference among the 14 tissues by RefFinder. Furthermore, the recommended comprehensive ranking of the stability determined by RefFinder showed that 18S was the most stable gene during the early developmental stages (from oosphere to 90 days old) in this study. However, when divided the C t value data of the above mentioned early developmental stages into two separate periods (embryo and post-hatching periods), TUBA and 18S represented the most stable references of these two developmental periods, respectively. Consequently, the reference gene should be carefully and accurately chosen even for studies of the same species at various developmental processes. The relevant data may help in selecting appropriate reference genes for mRNA expression analysis, and is of great value in the studies of fish growth and development.

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Authors and Affiliations

  1. Key Laboratory of Sustainable Development of Marine Fisheries of Ministry of Agriculture, Shandong Provincial Key Laboratory of Fishery Resources and Eco-environment, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071, China

    Qian Ma, Zhimeng Zhuang, Wenrong Feng, Shufang Liu & Qisheng Tang

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  1. Qian Ma
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  2. Zhimeng Zhuang
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  3. Wenrong Feng
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  4. Shufang Liu
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  5. Qisheng Tang
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Correspondence to Zhimeng Zhuang.

Additional information

Foundation item: The National Natural Science Foundation of China under contract No. 31201981; China Postdoctoral Science Foundation under contract No. 2013M531658; the Special Scientific Research Funds for Central Non-profit Institutes, Yellow Sea Fisheries Research Institutes under contract No. 20603022012032.

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Ma, Q., Zhuang, Z., Feng, W. et al. Evaluation of reference genes for quantitative real-time PCR analysis of gene expression during early development processes of the tongue sole (Cynoglossus semilaevis). Acta Oceanol. Sin. 34, 90–97 (2015). https://doi.org/10.1007/s13131-015-0725-5

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  • DOI: https://doi.org/10.1007/s13131-015-0725-5

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