Nature Biotechnology
17, 798 - 803 (1999)
doi:10.1038/11743
Gene expression analysis by transcript profiling coupled to a gene database
queryRichard A. Shimkets1, 4, David G. Lowe2, 4, Julie Tsu-Ning Tai2, 3, Patricia Sehl3, Hongkui Jin2, Renhui Yang2, Paul F. Predki1, Bonnie E. G. Rothberg1, Michael T. Murtha1, Matthew E. Roth1, Suresh G. Shenoy1, Andreas Windemuth1, John W. Simpson1, Jan F. Simons1, Michael P. Daley1, Steven A. Gold1, Michael P. McKenna1, Kenneth Hillan2, Gregory T. Went1
& Jonathan M. Rothberg11
CuraGen Corporation, 555 Long Wharf Drive
, New Haven, CT 06511. 2
Departments of Cardiovascular Research and Pathology,
Genentech, Inc., 1 DNA Way, South San Francisco
, CA 94080. 3
Current address: CVTherapeutics, 3172
Porter Dr., Palo Alto, CA 94304. 4
Both authors contributed equally to this work.
Correspondence should be addressed to Richard A. Shimkets Rshimket@curagen.comcardiac hypertrophydifferential gene expressiontranscript profilingoligonucleotide poisoning1We describe an mRNA profiling technique for determining differential gene
expression that utilizes, but does not require, prior knowledge of gene sequences.
This method permits high-throughput reproducible detection of most expressed
sequences with a sensitivity of greater than 1 part in 100,000. Gene identification
by database query of a restriction endonuclease fingerprint, confirmed by
competitive PCR using gene-specific oligonucleotides, facilitates gene discovery
by minimizing isolation procedures. This process, called GeneCalling, was
validated by analysis of the gene expression profiles of normal and hypertrophic
rat hearts following in vivo pressure overload.
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