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Selection of reference genes for RT-qPCR analysis in developing chicken embryonic ovary.
Wang, Yi; Zhang, Yu-Qing; Wu, Zi-Wei; Fang, Ting; Wang, Fang; Zhao, Han; Du, Zhi-Qiang; Yang, Cai-Xia.
Afiliación
  • Wang Y; College of Animal Science, Yangtze University, 434025, Jingzhou, Hubei, China.
  • Zhang YQ; College of Animal Science, Yangtze University, 434025, Jingzhou, Hubei, China.
  • Wu ZW; College of Animal Science, Yangtze University, 434025, Jingzhou, Hubei, China.
  • Fang T; College of Animal Science, Yangtze University, 434025, Jingzhou, Hubei, China.
  • Wang F; College of Animal Science, Yangtze University, 434025, Jingzhou, Hubei, China.
  • Zhao H; College of Animal Science, Yangtze University, 434025, Jingzhou, Hubei, China.
  • Du ZQ; College of Animal Science, Yangtze University, 434025, Jingzhou, Hubei, China. zhqdu@yangtzeu.edu.cn.
  • Yang CX; College of Animal Science, Yangtze University, 434025, Jingzhou, Hubei, China. caixiayang@yangtzeu.edu.cn.
Mol Biol Rep ; 50(4): 3379-3387, 2023 Apr.
Article en En | MEDLINE | ID: mdl-36729208
BACKGROUND: Normalization of the expression profiling of target genes, in a tissue-specific manner and under different experimental conditions, requires stably expressed gene(s) to be used as internal reference(s). However, to study the molecular regulation of oocyte meiosis initiation during ovary development in chicken embryos, stable reference gene(s) still need to be compared and confirmed. METHODS AND RESULTS: Six candidate genes previously used as internal references for the chicken embryo (Actb, Cvh, Dazl, Eef1a, Gapdh and Rpl15) were chosen, and their expression profiles in left ovaries dissected at five chicken embryonic days (E12.5, E15.5, E17.5, E18.5 and E20.5) were evaluated, respectively. Separately, GeNorm, NormFinder, BestKeeper and Comparative ΔCt methods were used to assess the stability of candidate reference genes, and all results were combined to give the final rank by RefFinder. All methods identified that Eef1a and Rpl15 were the two most stable internal reference genes, whereas Cvh is the most unstable one. Moreover, expression levels of three marker genes for chicken oocyte meiosis entry (Stra8, Scp3 and Dmc1) were normalized, based on Eef1a, Rpl15, or their combinations, respectively. CONCLUSION: Our findings provide the most suitable internal reference genes (Eef1a and Rpl15), to investigate further molecular regulation of ovary development and oocyte meiosis initiation in chicken embryos.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Pollos / Perfilación de la Expresión Génica Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Mol Biol Rep Año: 2023 Tipo del documento: Article País de afiliación: China Pais de publicación: Países Bajos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Pollos / Perfilación de la Expresión Génica Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Mol Biol Rep Año: 2023 Tipo del documento: Article País de afiliación: China Pais de publicación: Países Bajos