Identification of suitable normalizing genes for quantitative real-time RT-PCR analysis of gene expression in fetal mouse gonads

University of Queensland Your Subscriptions Logo Vol. 3, No. 4, 2009 Free Abstract Article (Fulltext) Article (PDF 395 KB) Original Article Identification of Suitable Normalizing Genes for Quantitative Real-Time RT-PCR Analysis of Gene Expression in Fetal Mouse Gonads T. Svingena, C.M. Spillera, K. Kashimadaa, V.R. Harleyb, P. Koopmana aDivision of Molecular Genetics and Development, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Qld., bHuman Molecular Genetics Laboratory, Prince Henry's Institute of Medical Research, Clayton, Vic., Australia Address of Corresponding Author Sex Dev 2009;3:194-204 (DOI: 10.1159/000228720) goto top of page Key Words * Housekeeping genes * Ovary * Polymerase chain reaction * Sexual development * Testis goto top of page Abstract In biological research, quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) assays are commonly employed to study mRNA abundance in cells and tissues. This type of assay usually relies on assessing transcript abundance relative to constitutively expressed endogenous reference genes. Therefore, it is important that the reference genes themselves are stably expressed in the cells or tissues analyzed, independent of factors such as age, sex, disease or experimental manipulations. Since no gene is expressed at the same level in all cells at all times, suitable reference genes must be identified for the specific cellular system or tissue being investigated. Here, we sought to identify stably expressed endogenous reference genes during embryonic gonad development in the mouse. We measured the transcript abundance of 10 frequently employed normalizing genes, of which 4 were stably expressed in fetal gonads from 11.5 to 14.5 dpc irrespective of sex. Based on our analysis, we suggest that Rn18s, Rps29, Tbp and Sdha are suitable reference genes for qRT-PCR expression studies during early gonad differentiation in the mouse