Tissue gene expression analysis using arrayed normalized cDNA libraries.
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.
Eickhoff
H
H
Max-Planck-Institut für Molekulare Genetik, 14195 Berlin, Germany. eickhoff@molgen.mpg.de
Schuchhardt
J
J
Ivanov
I
I
Meier-Ewert
S
S
O'Brien
J
J
Malik
A
A
Tandon
N
N
Wolski
E W
EW
Rohlfs
E
E
Nyarsik
L
L
Reinhardt
R
R
Nietfeld
W
W
Lehrach
H
H
D000465:Algorithms; D000818:Animals; D016172:DNA Fingerprinting / Q000295:instrumentation / Q000379:methods; D005333:Fetus / Q000737:chemistry; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods*; D015723:Gene Library; D051379:Mice; D008969:Molecular Sequence Data; D005810:Multigene Family; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D010802:Phylogeny; D051381:Rats