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dc.contributor.authorBagés, S.
dc.contributor.authorEstany Illa, Joan
dc.contributor.authorTor i Naudí, Marc
dc.contributor.authorPena i Subirà, Ramona Natacha
dc.date.accessioned2018-05-29T12:22:31Z
dc.date.available2018-05-29T12:22:31Z
dc.date.issued2015-02-11
dc.identifier.issn0378-1119
dc.identifier.urihttp://hdl.handle.net/10459.1/63449
dc.description.abstractAccurate normalization of data is required to correct for different efficiencies and errors during the processing of samples in reverse transcription PCR analysis. The chicken is one of the main livestock species and its genome was one of the first reported and used in large scale transcriptomic analysis. Despite this, the chicken has not been investigated regarding the identification of reference genes suitable for the quantitative PCR analysis of growth and fattening genes. In this study, five candidate reference genes (B2M, RPL32, SDHA, TBP and YWHAZ) were evaluated to determine the most stable internal reference for quantitative PCR normalization in the two main commercial muscles (pectoralis major (breast) and biceps femoris (thigh)), liver and abdominal fat. Four statistical methods (geNorm, NormFinder, CV and BestKeeper) were used in the evaluation of the most suitable combination of reference genes. Additionally, a comprehensive ranking was established with the RefFinder tool. This analysis identified YWHAZ and TBP as the recommended combination for the analysis of biceps femoris and liver, YWHAZ and RPL32 for pectoralis major and RPL32 and B2M for abdominal fat and across-tissue studies. The final ranking for each tool changed slightly but overall the results, and most particularly the ability to discard the least robust candidates, were consistent between tools. The selection and number of reference genes were validated using SCD, a target gene related to fat metabolism. Overall, the results can be directly used to quantitate target gene expression in different tissues or in validation studies from larger transcriptomic experiments.
dc.description.sponsorshipS. Bagés received a Research Starting Grant from the University of Lleida (47900461V). This work is partially supported by the Spanish Ministry of Economy and Competitiveness (MINECO; grants AGL2012- 33529 and PIM2010PKB-00746).
dc.format.mimetypeapplication/pdf
dc.language.isoeng
dc.publisherElsevier
dc.relationMICINN/PN2008-2011/AGL2012-33529
dc.relationMICINN/PN2008-2011/PIM2010PKB-00746
dc.relation.isformatofVersió postprint del document publicat a: https://doi.org/10.1016/j.gene.2015.02.016
dc.relation.ispartofGene, 2015, vol. 561, num. 1, p. 82-87
dc.rightscc-by-nc-nd (c) Elsevier, 2015
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectqRT-PCR
dc.subjectGene expression
dc.subjectNormalization
dc.subjectEndogenous control
dc.subject.otherAnimals--Cria i desenvolupament
dc.subject.otherGenètica animal
dc.subject.otherGenètica molecular
dc.titleInvestigating reference genes for quantitative real-time PCR analysis across four chicken tissues
dc.typeinfo:eu-repo/semantics/article
dc.date.updated2018-05-29T12:22:31Z
dc.identifier.idgrec022326
dc.type.versioninfo:eu-repo/semantics/acceptedVersion
dc.rights.accessRightsinfo:eu-repo/semantics/openAccess
dc.identifier.doihttps://doi.org/10.1016/j.gene.2015.02.016


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cc-by-nc-nd (c) Elsevier, 2015
Except where otherwise noted, this item's license is described as cc-by-nc-nd (c) Elsevier, 2015