Gene expression analysis by transcript profiling coupled to a gene database query

We 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. Ge...

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Bibliographic Details
Published in:Nature biotechnology 1999-08, Vol.17 (8), p.798-803
Main Authors: Shimkets, Richard A, Lowe, David G, Tai, Julie Tsu-Ning, Sehl, Patricia, Jin, Hongkui, Yang, Renhui, Predki, Paul F, Rothberg, Bonnie E. G, Murtha, Michael T, Roth, Matthew E, Shenoy, Suresh G, Windemuth, Andreas, Simpson, John W, Simons, Jan F, Daley, Michael P, Gold, Steven A, McKenna, Michael P, Hillan, Kenneth, Went, Gregory T, Rothberg, Jonathan M
Format: Article
Language:English
Subjects:
Agriculture
Animals
Bioinformatics
Biological and medical sciences
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
Cardiomegaly - genetics
Cardiomegaly - pathology
Databases, Factual
Diverse techniques
Fundamental and applied biological sciences. Psychology
Gene Expression
HeLa Cells
Humans
Life Sciences
Male
Molecular and cellular biology
Rats
Rats, Sprague-Dawley
Reproducibility of Results
research-article
RNA, Messenger - genetics
Sensitivity and Specificity
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Description
Summary:We 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.
ISSN:1087-0156
1546-1696
DOI:10.1038/11743