International Mammalian Genome Society

The 15th International Mouse Genome Conference (2001)


POSTER 113 - MOUSEEXPRESS: DNA-MICROARRAYS AND REPRESENTATIONAL DIFFERENCE ANALYSIS (RDA) FOR GLOBAL TRANSCRIPTIONAL PROFILING OF ENU-INDUCED MOUSE MUTANTS

Ms Tamara Korica
German Cancer Research Centre
Im Neuenheimer Feld 506
Heidelberg
69120
Germany

Co-Authors: 1)Koenig R, 1)Frohme M, 2)Drobyshev A, 2)Seltmann M, 2)Chen Y, 2)Machka C, 3)Mader M, 3)Tornow S, 2)Hrabe de Angelis M, 3)Mewes W, 4)Vingron M, 2)Beckers J, 1)Hoheisel J.
Institutions: 1)DKFZ, Division of Functional Genome Analysis, 2)GSF - National Research Center for Environment and Health, Institute of Experimental Genetics, 3)GSF - National Research Center for Environment and Health, Münchener Informationszentrum für Proteinsequenzen (MIPS), 4)MPI für Molekulare Genetik, Computational Molecular Biology,

As part of German Human Genome Project (DHGP) we perform a systematic transcriptome analysis of ENU induced mouse mutant lines (GSF/LMU, Munich). The MouseExpress project involves specialists in mouse genetics, microarray technologies and bioinformatics. For transcriptional profiling used are 17 different tissues and organs from 300 mouse mutant strains, produced in on-going ENU mutagenesis screening project.

A genome-wide, non-redundant EST clone set has been selected by the well established cluster and assembly analysis software of the Gene Nest project (TBI-DKFZ, Heidelberg). Clones are selected from the IMAGE consortium library and provided by RZPD, Berlin.

For the initial analysis, we assembled a gene function specific DNA-chip containing 400 genes in a library of 1200 EST-clones, thus producing general calibration and normalization data-sets. Experimental variables included, for example, different slide surfaces, spotting conditions and labelling methods. We have developed protocols, and currently are monitoring the performance of our own set of external controls, involving spiking poly-A tailed ssDNA instead of RNA, gaining thereby reliability. As an additional method for the investigation of differential gene expression, we use representational difference analysis of cDNA. The outcome of several experiments was used to complement the library analysed on gene-specific array.


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