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Effect of Embryonic Genotype on Reference Gene Selection for RT-qPCR Normalization

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Effect of Embryonic Genotype on Reference Gene Selection for RT-qPCR Normalization

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dc.contributor.author Llobat, L. es_ES
dc.contributor.author Marco Jiménez, Francisco es_ES
dc.contributor.author Peñaranda, D.S. es_ES
dc.contributor.author Saenz de Juano Ribes, María de los Desamparados es_ES
dc.contributor.author Vicente Antón, José Salvador es_ES
dc.date.accessioned 2017-05-10T10:34:42Z
dc.date.available 2017-05-10T10:34:42Z
dc.date.issued 2012-08
dc.identifier.issn 0936-6768
dc.identifier.uri http://hdl.handle.net/10251/80810
dc.description.abstract [EN] Contents To obtain more reliable results with biological significance, it requires data normalization using an appropriate internal control gene. In rabbits, there are classic stable reference genes that have been identified for normalization in oocytes and pre-implantation stage embryos. However, effects of embryonic genotype on reference gene selection have not been elucidated. The aim of this study was to test (i) the stability of mRNA transcription level for histone (H2afz) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) genes in rabbit blastocysts from two lines selected by different criteria (litter size and post-weaning daily weight gain) and (ii) its influence on biological significance examined by means of a set of embryonic transcripts, such as POU5F1 (Oct-4), epidermal growth factor receptor (erbB3), transforming growth factor-beta2, vascular endothelial growth factor and gamma interferon (Ifn-gamma). The geNorm, NormFinder and BestKeeper programs showed similar results, pointing out that H2afz and GAPDH were the most stable reference genes in rabbits selected on litter size at weaning. Moreover, our study revealed that embryonic genotype affected target gene expression when a single reference gene was used to analyse mRNA expression in blastocysts. Results showed that GAPDH gene is better than H2afz for gene expression studies of both embryo genotypes. A normalization factor derived from H2afz and GAPDH is likely to be appropriate when RT-qPCR was performed in rabbit embryos with different genotypes. es_ES
dc.description.sponsorship We are grateful to the Researcher Personal Training Programme from Valencia Polytechnic University. This work was supported by the Spanish Research Project (CICYT AGL2008-03274) and Generalitat Valenciana Research Programme (Prometeo 2009/125). The authors thank Neil Macowan Language Services for revising the English version of the manuscript. en_EN
dc.language Inglés es_ES
dc.publisher Wiley es_ES
dc.relation.ispartof Reproduction in Domestic Animals es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject Bovine preimplantation embryos es_ES
dc.subject Transcription PCR data es_ES
dc.subject Time quantitative PCR es_ES
dc.subject Real time es_ES
dc.subject Housekeeping genes es_ES
dc.subject In vitro es_ES
dc.subject Relative quantification es_ES
dc.subject Expression es_ES
dc.subject Validation es_ES
dc.subject Oocytes es_ES
dc.subject.classification BIOLOGIA ANIMAL es_ES
dc.subject.classification PRODUCCION ANIMAL es_ES
dc.title Effect of Embryonic Genotype on Reference Gene Selection for RT-qPCR Normalization es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1111/j.1439-0531.2011.01934.x
dc.relation.projectID info:eu-repo/grantAgreement/MICINN//AGL2008-03274/ES/MEJORA GENETICA DEL CONEJO DE CARNE: REPRODUCCION, ESTRES Y LONGEVIDAD/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/GVA//PROMETEO%2F2009%2F125/ES/Efecto de la crioconservación de embriones sobre el desarrollo y el re-establecimiento de poblaciones/ es_ES
dc.rights.accessRights Cerrado es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal es_ES
dc.contributor.affiliation Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal es_ES
dc.description.bibliographicCitation Llobat, L.; Marco Jiménez, F.; Peñaranda, D.; Saenz De Juano Ribes, MDLD.; Vicente Antón, JS. (2012). Effect of Embryonic Genotype on Reference Gene Selection for RT-qPCR Normalization. Reproduction in Domestic Animals. 47(4):629-634. https://doi.org/10.1111/j.1439-0531.2011.01934.x es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion http://doi.org/10.1111/j.1439-0531.2011.01934.x es_ES
dc.description.upvformatpinicio 629 es_ES
dc.description.upvformatpfin 634 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 47 es_ES
dc.description.issue 4 es_ES
dc.relation.senia 205376 es_ES
dc.identifier.pmid 22044783
dc.contributor.funder Ministerio de Ciencia e Innovación es_ES
dc.contributor.funder Generalitat Valenciana es_ES


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