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• W1987608719 endingPage "e99678" @default.
• W1987608719 startingPage "e99678" @default.
• W1987608719 abstract "Quantitative real-time PCR (qPCR) rapidly and reliably quantifies gene expression levels across different experimental conditions. Selection of suitable reference genes is essential for meaningful normalization and thus correct interpretation of data. In recent years, an increasing number of avian species other than the chicken has been investigated molecularly, highlighting the need for an experimentally validated pan-avian primer set for reference genes. Here we report testing a set for 14 candidate reference genes (18S, ABL, GAPDH, GUSB, HMBS, HPRT, PGK1, RPL13, RPL19, RPS7, SDHA, TFRC, VIM, YWHAZ) on different tissues of the mallard (Anas platyrhynchos), domestic chicken (Gallus gallus domesticus), common crane (Grus grus), white-tailed eagle (Haliaeetus albicilla), domestic turkey (Meleagris gallopavo f. domestica), cockatiel (Nymphicus hollandicus), Humboldt penguin (Sphenicus humboldti), ostrich (Struthio camelus) and zebra finch (Taeniopygia guttata), spanning a broad range of the phylogenetic tree of birds. Primer pairs for six to 11 genes were successfully established for each of the nine species. As a proof of principle, we analyzed expression levels of 10 candidate reference genes as well as FOXP2 and the immediate early genes, EGR1 and CFOS, known to be rapidly induced by singing in the avian basal ganglia. We extracted RNA from microbiopsies of the striatal song nucleus Area X of adult male zebra finches after they had sang or remained silent. Using three different statistical algorithms, we identified five genes (18S, PGK1, RPS7, TFRC, YWHAZ) that were stably expressed within each group and also between the singing and silent conditions, establishing them as suitable reference genes. In conclusion, the newly developed pan-avian primer set allows accurate normalization and quantification of gene expression levels in multiple avian species." @default.
• W1987608719 created "2016-06-24" @default.
• W1987608719 creator A5011653179 @default.
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• W1987608719 creator A5054933257 @default.
• W1987608719 date "2014-06-13" @default.
• W1987608719 modified "2023-09-30" @default.
• W1987608719 title "Reference Genes for Quantitative Gene Expression Studies in Multiple Avian Species" @default.
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• W1987608719 doi "https://doi.org/10.1371/journal.pone.0099678" @default.
• W1987608719 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4057121" @default.
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• W1987608719 hasPublicationYear "2014" @default.
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