Evaluation of reference genes for real-time RT-PCR expression studies in the plant pathogen Pectobacterium atrosepticum.docVIP

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Evaluation of reference genes for real-time RT-PCR expression studies in the plant pathogen Pectobacterium atrosepticum.doc

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Evaluation of reference genes for real-time RT-PCR expression studies in the plant pathogen Pectobacterium atrosepticum

BMC Plant Biology BioMedCentral Research article Open Access Evaluation of reference genes for real-time RT-PCR expression studies in the plant pathogen Pectobacterium atrosepticum Gunnhild W Takle1,2, Ian K Toth3 and May B Brurberg*1 Address: 1Norwegian Institute for Agricultural and Environmental Research, Plant Health and Plant Protection Division, H?gskoleveien 7, 1432 ?s, Norway, 2Norwegian University of Life Sciences, Institute for Chemistry, Biotechnology and Food Science, PO Box 5003, 1432 ?s, Norway and 3SCRI, Invergowrie, Dundee DD2 5DA, UK Email: Gunnhild W Takle - gunnhild.takle@bioforsk.no; Ian K Toth - ian.toth@scri.ac.uk; May B Brurberg* - may.brurberg@bioforsk.no * Corresponding author Published: 21 September 2007 Received: 13 February 2007 Accepted: 21 September 2007 BMC Plant Biology 2007, 7:50 doi:10.1186/1471-2229-7-50 This article is available from: /1471-2229/7/50 ? 2007 Takle 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 RT-PCR has become a powerful technique to monitor low-abundance mRNA expression and is a useful tool when examining bacterial gene expression inside infected host tissues. However, correct evaluation of data requires accurate and reliable normalisation against internal standards. Thus, the identification of reference genes whose expression does not change during the course of the experiment is of paramount importance. Here, we present a study where manipulation of cultural growth conditions and in planta experiments have been used to validate the expression stability of reference gene candidates for the plant pathogen Pectobacterium atrosepticum, belonging to the family Enterobacteriaceae . Results: Of twelve refe