A GeNorm algorithm-based selection of reference genes for quantitative real-time PCR in skin biopsies of healthy dogs and dogs with atopic dermatitis

dc.contributor.authorSchlotter, Yvette M.
dc.contributor.authorVeenhof, Eveline Z.
dc.contributor.authorBrinkhof, Bas
dc.contributor.authorRutten, Victor P.M.G.
dc.contributor.authorSpee, Bart
dc.contributor.authorWillemse, Ton
dc.contributor.authorPenning, Louis C.
dc.date.accessioned2009-04-07T10:51:53Z
dc.date.available2009-04-07T10:51:53Z
dc.date.issued2009
dc.description.abstractQuantitative real time PCR (Q-PCR) is the method of choice to study mRNA expression levels. Since Q-PCR is very sensitive, normalization of the data with stably expressed reference genes if of utmost importance. The stability of reference genes depends on the tissue and the species of interest. Therefore, evaluation of the stability of reference genes must be performed for each new tissue and species under study. The stability of B2M, GAPDH, HPRT, SRPR, hnRNPH, GUSB, RPL8, RPS5, and RPS19 was analyzed with the GeNorm software in snap frozen canine skin biopsies. Healthy dogs (n = 7) and dogs with confirmed atopic dermatitis (n = 28) were included. Lesional and non-lesional skin was analyzed. The study indicated that themost appropriate reference genes in canine skin are the ribosomal gene products RPL8, RPS5 and RPS19 besides GUSB and HPRT. As little as three reference genes will reveal highly reliable Q-PCR calculations.en
dc.identifier.citationSchlotter, Y.M., et al., A GeNorm algorithm-based selection of reference genes for quantitative real-time PCR in skin biopsies of healthy dogs and dogs with atopic dermatitis. Veterinary Immunology and Immunopathology (2009), doi:10.1016/j.vetimm.2008.12.004en
dc.identifier.issn0165-2427
dc.identifier.other10.1016/j.vetimm.2008.12.004
dc.identifier.urihttp://hdl.handle.net/2263/9497
dc.language.isoenen
dc.publisherElsevieren
dc.relation.requiresAdobe Acrobat Readeren
dc.rightsElsevieren
dc.subjectReference genesen
dc.subjectCanine skinen
dc.subjectAtopic dermatitis (AD)
dc.subject.lcshGenesen
dc.subject.lcshPolymerase chain reaction (PCR)en
dc.subject.lcshDogs -- Diseasesen
dc.subject.lcshAllergyen
dc.subject.lcshSkin -- Inflammationen
dc.titleA GeNorm algorithm-based selection of reference genes for quantitative real-time PCR in skin biopsies of healthy dogs and dogs with atopic dermatitisen
dc.typePostprint Articleen

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