Biotechnology Letters

, Volume 26, Issue 6, pp 509–515 | Cite as

Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper – Excel-based tool using pair-wise correlations

  • Michael W. Pfaffl
  • Ales Tichopad
  • Christian Prgomet
  • Tanja P. Neuvians


The stability of standard gene expression is an elementary prerequisite for internal standardisation of target gene expression data and many so called housekeeping genes with assumed stable expression can exhibit either up- or down-regulation under some experimental conditions. The developed, and herein presented, software called BestKeeper determines the best suited standards, out of ten candidates, and combines them into an index. The index can be compared with further ten target genes to decide, whether they are differentially expressed under an applied treatment. All data processing is based on crossing points. The BestKeeper software tool was validated on four housekeeping genes and 10 members of the somatotropic axis differentially expressed in bovine corpora lutea total RNA. The BestKeeper application and necessary information about data processing and handling can be downloaded on

β-actin housekeeping gene PCR normalisation RT-PCR somatotropic axis ubiquitin 


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Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Michael W. Pfaffl
    • 1
  • Ales Tichopad
    • 1
  • Christian Prgomet
    • 1
  • Tanja P. Neuvians
    • 1
  1. 1.Physiology, FML-Weihenstephan, Centre of Life and Food ScienceTechnical University of MunichGermany

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