Validating internal controls for quantitative plant gene expression studies 英文参考文献.docVIP

BMC Plant Biology BioMedCentral Methodology article Open Access Validating internal controls for quantitative plant gene expression studies Amy M Brunner*?1, Igor A Yakovlev?1,2 and Steven H Strauss1 Address: 1Department of Forest Science, Oregon State University, Corvallis, OR 97331-5752, USA and 2Skogforsk/ Norwegian Forest Research Institute, Hogskoleveien 12, N-1432 As, Norway Email: Amy M Brunner* - Amy.Brunner@; Igor A Yakovlev - Igor.Yakovlev@skogforsk.no; Steven H Strauss - Steve.Strauss@ * Corresponding author ?Equal contributors Published: 18 August 2004 Received: 20 December 2003 Accepted: 18 August 2004 BMC Plant Biology 2004, 4:14 doi:10.1186/1471-2229-4-14 This article is available from: /1471-2229/4/14 ? 2004 Brunner et al; licensee BioMed Central Ltd. This is an open-access article distributed under the terms of the Creative Commons Attribution License (/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: Real-time reverse transcription PCR (RT-PCR) has greatly improved the ease and sensitivity of quantitative gene expression studies. However, accurate measurement of gene expression with this method relies on the choice of a valid reference for data normalization. Studies rarely verify that gene expression levels for reference genes are adequately consistent among the samples used, nor compare alternative genes to assess which are most reliable for the experimental conditions analyzed. Results: Using real-time RT-PCR to study the expression of 10 poplar (genus Populus) housekeeping genes, we demonstrate a simple method for determining the degree of stability of gene expression over a set of experimental conditions. Based on a traditional method for analyzing the stability of varieties in plant breeding, it defines measures of gene expression stability from analysis of variance