Selection of reference genes for quantitative RT-PCR (RT-qPCR) analysis of rat tissues under physiological and toxicological conditions

• DOI: 10.7717/peerj.855
• Publication Date: 2015-03-24T04:43:46Z
• Authors: Terje Svingen; Heidi Letting; Niels Hadrup; Ulla Hass; Anne Marie Vinggaard
• Keywords: Reference Genes; SDHA; Housekeeping gene; Toxicogenomics; Glyceraldehyde 3-phosphate dehydrogenase

In biological research the analysis of gene expression levels in cells and tissues can be a powerful tool to gain insights into processes. For this, quantitative RT-PCR (RT-qPCR) is popular method that often involve use constitutively expressed endogenous reference (or 'housekeeping') for normalization data. Thus, it essential genes have been verified stably within specific experimental setting. Here, we analysed stability 12 commonly used (Actb, B2m, Gapdh, Hprt, Pgk1, Rn18s, Rpl13a, Rps18, Rps29, Sdha, Tbp Ubc) across several juvenile adult rat (liver, adrenal, prostate, fat pad, testis ovaries), both under normal conditions following exposure various chemicals during development. Employing NormFinder BestKeeper softwares, found Hprt Sdha amongst most stable manipulated tissues, with others also being suitable tissues. B2m displayed highest variability transcript between developmental stages. It was observed were unstable liver toxicological exposure. future studies, propose more than one continuous monitoring their suitability conditions, including based on changes threshold (Ct) values from cDNA samples having reverse-transcribed constant input concentration RNA.