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. 2007 Nov 15;39(6):633–650. doi: 10.1186/1297-9686-39-6-633

Analysis of the real EADGENE data set: Comparison of methods and guidelines for data normalisation and selection of differentially expressed genes (Open Access publication)

Florence Jaffrézic 1,, Dirk-Jan de Koning 2, Paul J Boettcher 3, Agnès Bonnet 4, Bart Buitenhuis 5, Rodrigue Closset 6, Sébastien Déjean 7, Céline Delmas 8, Johanne C Detilleux 9, Peter Dovč 10, Mylène Duval 8, Jean-Louis Foulley 1, Jakob Hedegaard 5, Henrik Hornshøj 5, Ina Hulsegge 11, Luc Janss 5, Kirsty Jensen 2, Li Jiang 5, Miha Lavrič 10, Kim-Anh Lê Cao 7,8, Mogens Sandø Lund 5, Roberto Malinverni 3, Guillemette Marot 1, Haisheng Nie 12, Wolfram Petzl 13, Marco H Pool 11, Christèle Robert-Granié 8, Magali San Cristobal 4, Evert M van Schothorst 14, Hans-Joachim Schuberth 15, Peter Sørensen 5, Alessandra Stella 3, Gwenola Tosser-Klopp 4, David Waddington 2, Michael Watson 16, Wei Yang 17, Holm Zerbe 13, Hans-Martin Seyfert 17
PMCID: PMC2682811  PMID: 18053573

Abstract

A large variety of methods has been proposed in the literature for microarray data analysis. The aim of this paper was to present techniques used by the EADGENE (European Animal Disease Genomics Network of Excellence) WP1.4 participants for data quality control, normalisation and statistical methods for the detection of differentially expressed genes in order to provide some more general data analysis guidelines. All the workshop participants were given a real data set obtained in an EADGENE funded microarray study looking at the gene expression changes following artificial infection with two different mastitis causing bacteria: Escherichia coli and Staphylococcus aureus. It was reassuring to see that most of the teams found the same main biological results. In fact, most of the differentially expressed genes were found for infection by E. coli between uninfected and 24 h challenged udder quarters. Very little transcriptional variation was observed for the bacteria S. aureus. Lists of differentially expressed genes found by the different research teams were, however, quite dependent on the method used, especially concerning the data quality control step. These analyses also emphasised a biological problem of cross-talk between infected and uninfected quarters which will have to be dealt with for further microarray studies.

Keywords: quality control, differentially expressed genes, mastitis resistance, microarray data, normalisation

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