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Combined subtractive cDNA cloning and array CGH an efficient approach for identification of overexpressed genes in DNA amplicons.doc

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BMC Genomics BioMedCentral Research article Open Access Combined subtractive cDNA cloning and array CGH: an efficient approach for identification of overexpressed genes in DNA amplicons Katleen De Preter1, Filip Pattyn1, Geert Berx2, Kristin Strumane2, Bj?rn Menten1, Frans Van Roy2, Anne De Paepe1, Frank Speleman1 and Jo Vandesompele* Address: 1Center for Medical Genetics, Ghent University Hospital 1K5, De Pintelaan 185, 9000 Gent, Belgium and 2Department for Molecular Biomedical Research, Flemish Interuniversity Institute for Biotechnology (VIB), Ghent University, Technologiepark 927, 9052 Zwijnaarde, Belgium Email: Katleen De Preter - katleen.depreter@ugent.be; Filip Pattyn - filip.pattyn@ugent.be; Geert Berx - geert.berx@dmbr.ugent.be; Kristin Strumane - kristin.strumane@dmbr.ugent.be; Bj?rn Menten - bjorn.menten@ugent.be; Frans Van Roy - frans.vanroy@dmbr.ugent.be; Anne De Paepe - anne.depaepe@ugent.be; Frank Speleman - franki.speleman@ugent.be; Jo Vandesompele* - joke.vandesompele@ugent.be * Corresponding author Published: 03 February 2004 Received: 03 October 2003 Accepted: 03 February 2004 BMC Genomics 2004, 5:11 This article is available from: /1471-2164/5/11 ? 2004 De Preter et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the articles original URL. AmplificationOverexpressionOncogeneSSHArray CGHNeuroblastoma Abstract Background: Activation of proto-oncogenes by DNA amplification is an important mechanism in the development and maintenance of cancer cells. Until recently, identification of the targeted genes relied on labour intensive and time consuming positional cloning methods. In this study, we outline a straightforward and efficient strategy for fast and comprehensive cloning of amplified and overexpressed genes. Results: As a proof of principle, we analyz