Screening of Stably Expressed Internal Reference Genes for Quantitative Real-Time PCR Analysis in Quail

Z. W. Yuan; X. H. Zhang; Y. Z. PangY. X. QiQ. K. WangS. W. RenY. Q. HuY. W. ZhaoT. WangL. K. Huo

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Screening of Stably Expressed Internal Reference Genes for Quantitative Real-Time PCR Analysis in Quail

机译：Screening of Stably Expressed Internal Reference Genes for Quantitative Real-Time PCR Analysis in Quail

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Abstract This experiment analyzed the expression levels of 10 genes in quail embryo tissues at different developmental stages, and screened the most stable internal reference genes (IRGs). Total RNA was extracted from quail whole embryonic tissue at 4 different developmental stages (8d, 10d, 12d and 14d) and reverse transcribed to cDNA. Using the microarray data (GSE28388) during chicken embryo development, 8 genes with moderate expression levels and the most stable expression during the development of chicken embryo were initially screened as candidate IRGs, and the commonly used GAPDH and ACTB genes were used as controls. Absolute quantitative RT-qPCR was used to analyze the expression changes of candidate reference genes during quail embryo development. The expression stability of these genes was analyzed using geNorm, NormFinder and BestKeeper program. The results of this experiment showed that the most suitable IRGs in quail at different developmental periods was EIF2S3, and the most suitable combination of IRGs was EIF2S3 and RPL10A. The stability of the IRGs screened in this experiment was verified by the analysis of MLANA gene expression. This provides a theoretical basis for the follow-up study of target gene expression of embryonic development in quail.

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《Biology bulletin》 |2022年第5期|418-427|共10页
作者
Z. W. Yuan; X. H. Zhang; Y. Z. PangY. X. QiQ. K. WangS. W. RenY. Q. HuY. W. ZhaoT. WangL. K. Huo;
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作者单位

College of Animal Science, Henan University of Science and Technology;

College of Animal Science, Henan University of Science and Technology||Luoyang Key Laboratory of Animal Genetics and Breeding;

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正文语种英语
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quail; internal reference gene; real-time quantitative PCR; expression stability;

Screening of Stably Expressed Internal Reference Genes for Quantitative Real-Time PCR Analysis in Quail

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