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Vol. 10, Issue 8, 1230-1240, August 2000

METHODS
Tissue Gene Expression Analysis Using Arrayed Normalized cDNA Libraries

Holger Eickhoff,1,4 Johannes Schuchhardt,2 Igor Ivanov,3 Sebastian Meier-Ewert,3 John O'Brien,1 Arif Malik,1 Neeraj Tandon,1 Eryk-Witold Wolski,1 Elke Rohlfs,1 Lajos Nyarsik,1 Richard Reinhardt,1 Wilfried Nietfeld,1 and Hans Lehrach1

1 Max-Planck-Institut für Molekulare Genetik, 14195 Berlin, Germany; 2 Institut für Theoretische Biologie, Humboldt Universität Berlin, 10115 Berlin, Germany; 3 GPC-Biotech AG, 82152 Martinsried/Munich, Germany

We have used oligonucleotide-fingerprinting data on 60,000 cDNA clones from two different mouse embryonic stages to establish a normalized cDNA clone set. The normalized set of 5,376 clones represents different clusters and therefore, in almost all cases, different genes. The inserts of the cDNA clones were amplified by PCR and spotted on glass slides. The resulting arrays were hybridized with mRNA probes prepared from six different adult mouse tissues. Expression profiles were analyzed by hierarchical clustering techniques. We have chosen radioactive detection because it combines robustness with sensitivity and allows the comparison of multiple normalized experiments. Sensitive detection combined with highly effective clustering algorithms allowed the identification of tissue-specific expression profiles and the detection of genes specifically expressed in the tissues investigated. The obtained results are publicly available (http://www.rzpd.de) and can be used by other researchers as a digital expression reference.

[The sequence data described in this paper have been submitted to the EMBL data library under accession nos. AL360374-AL36537.]

4 Corresponding author.

10:1230-1240 ©2000 by Cold Spring Harbor Laboratory Press  ISSN 1088-9051/00 $5.00
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