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dc.contributor.authorMitter, Karin-
dc.contributor.authorKotoulas, Georgios-
dc.contributor.authorMagoulas, Antonios-
dc.contributor.authorMulero, Victoriano-
dc.contributor.authorSepulcre, Pilar-
dc.contributor.authorFigueras Huerta, Antonio-
dc.contributor.authorNovoa, Beatriz-
dc.contributor.authorSarropoulou, Elena-
dc.identifier.citationComparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 153(4): 340-347 (2009)en_US
dc.description8 pages, 6 figures, 5 tables.-- PMID: 19398033 [PubMed].-- Printed version published Aug 2009.en_US
dc.descriptionCorrigendum published in Comp Biochem Physiol B Biochem Mol Biol. Aug 6, 2009, http://dx.doi.org/10.1016/j.cbpb.2009.07.008-
dc.description.abstractThe expression level of mRNA can vary significantly in different experimental conditions, such as stress, infection, developmental stage or tissue. Suitable reference genes are expected to exhibit constant expression levels. However no single gene is constitutively expressed in all cell types and under all experimental conditions. It has become clear that expression stability of the intended reference gene has to be examined before each experiment. For expression studies using quantitative real-time PCR (qPCR) at least two reference genes have to be applied. So far expression studies in the European seabass (Dicentrarchus labrax) as well as in the Gilthead seabream (Sparus aurata) have been performed with only one reference gene (S18, Ef-1 alpha or Gapdh). Though significant variations showed up in other teleost species such as the Atlantic halibut and the zebrafish affirming the need for proper normalization strategies, the present study aims at identifying suitable reference genes among nine candidates [glyceraldehyde-phosphate-dehydrogenase (Gapdh), β-actin (two regions of β-actin), 40S ribosomal protein S30 (Fau), ribosomal protein L13 a (L13a), β2-tubulin (Tubb2) and tyrosine 3 monooxygenase/tryptophan 5-monooxygenase activation protein (Tyr)] for expression analysis of 8 developmental stages and a tissue panel (spleen, liver, kidney and brain) with samples infected with Nodavirus and Vibrio anguillarum in D. labrax. Besides the analysis of raw Ct-values, the gene expression stability was determined using two different software applications BestKeeper and NormFinder. According to both algorithms the best two reference genes for an appropriate normalization approach during D. labrax development are Ef-1 alpha and L13a whereas in the tissue panel Fau and L13a are recommended for qPCR normalization.en_US
dc.description.sponsorshipAuthors would like to thank the Institute of Aquaculture and specifically Dr. Nikos Papandroulakis and Constantinos Mylonas for providing embryonic and larvae stages. We also would like to thank Spiros Kollias for providing RNA.en_US
dc.format.extent918459 bytes-
dc.subjectReference geneen_US
dc.subjectDicentrarchus labraxen_US
dc.subjectDevelopmental stageen_US
dc.subjectVibrio anguillarumen_US
dc.titleEvaluation of candidate reference genes for QPCR during ontogenesis and of immune-relevant tissues of European seabass (Dicentrarchus labrax)en_US
dc.description.peerreviewedPeer revieweden_US
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