Identification of Suitable Normalizing Genes for Quantitative Real-Time RT-PCR Analysis of Gene Expression in Fetal Mouse Gonads

Terje Svingen, C. M. Spiller, K. Kashimada, V. R. Harley, P. Koopman

Research output: Contribution to journalJournal articleResearchpeer-review

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. Copyright (C) 2009 S. Karger AG, Basel
Original languageEnglish
JournalSexual Development
Volume3
Issue number4
Pages (from-to)194-204
Number of pages11
ISSN1661-5425
DOIs
Publication statusPublished - 2009
Externally publishedYes

Keywords

  • Animals
  • DNA Primers
  • Electrophoresis, Agar Gel
  • Fetus
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental
  • Gonads
  • Mesonephros
  • Mice
  • RNA, Messenger
  • Reference Standards
  • Reverse Transcriptase Polymerase Chain Reaction
  • animal experiment
  • article
  • cell differentiation
  • controlled study
  • embryo
  • embryo development
  • gene expression
  • genetic analysis
  • genetic identification
  • gonad development
  • mouse
  • nonhuman
  • priority journal
  • quantitative analysis
  • real time polymerase chain reaction
  • reverse transcription polymerase chain reaction
  • sexual development
  • Housekeeping genes
  • Ovary
  • Polymerase chain reaction
  • Sexual development
  • Testis
  • DEVELOPMENTAL
  • POLYMERASE CHAIN-REACTION
  • HOUSEKEEPING GENES
  • GERM-CELLS
  • QUANTIFICATION
  • SELECTION
  • TISSUE
  • RNA
  • INAPPROPRIATE
  • VALIDATION
  • STAGE
  • Rodentia Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Mammals, Nonhuman Vertebrates, Nonhuman Mammals, Rodents, Vertebrates) - Muridae [86375] mouse common fetus female
  • mouse Actb gene [Muridae] mouse beta-actin gene expression, reference gene
  • mouse Gapdh gene [Muridae] mouse glyceraldehyde-3 phosphate dehydrogenase gene expression, reference gene
  • mouse Hprt1 gene [Muridae] mouse hypoxanthine guanine phosphoribosyl transferase 1 gene expression, reference gene
  • mouse Pfkp gene [Muridae] mouse phosphofructokinase-platelet gene expression, reference gene
  • mouse Rn18s gene [Muridae] mouse 18S rRNA gene expression, reference gene
  • mouse Rps29 gene [Muridae] mouse ribosomal protein S29 gene expression, reference gene
  • mouse Sdha gene [Muridae] mouse succinate dehydrogenase complex-subunit A-flavoprotein gene expression, reference gene
  • mouse Tbp gene [Muridae] mouse TATA-box binding protein gene expression, reference gene
  • mouse Tuba1a gene [Muridae] mouse alpha-1a tubulin gene expression, reference gene
  • mouse Vim gene [Muridae] mouse vimentin gene expression, reference gene
  • transcript
  • 03502, Genetics - General
  • 03506, Genetics - Animal
  • 16504, Reproductive system - Physiology and biochemistry
  • 17002, Endocrine - General
  • 25502, Development and Embryology - General and descriptive
  • Endocrine System
  • Methods and Techniques
  • Molecular Genetics
  • Reproductive System
  • Biochemistry and Molecular Biophysics
  • Chemical Coordination and Homeostasis
  • Reproduction
  • gonad reproductive system, endocrine system
  • quantitative real-time reverse transcriptase-polymerase chain reaction qRT-PCR laboratory techniques, genetic techniques

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