ArticleNucleic acids research1998
Detection of differentially expressed genes in primary tumor tissues using representational differences analysis coupled to microarray hybridization.
Article in Nucleic acids research, 1998. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 172 citations in OpenAlex.
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- Molecular analysis of metastasis in a polyomavirus middle T mouse model: the role of osteopontin.Breast cancer research : BCR · 2004Article
- Modulation of GdCl3 and Angelica sinensis polysaccharides on differentially expressed genes in liver of hepatic immunological injury mice by cDNA microarray.World journal of gastroenterology · 2003Article
- Novel candidate tumor marker genes for lung adenocarcinoma.Oncogene · 2002Article
- Manufacturing DNA microarrays from unpurified PCR products.Nucleic acids research · 2002Article
- Comprehensive gene expression analysis by transcript profiling.Plant molecular biology · 2002Review
- DNA microarrays for functional plant genomics.Plant molecular biology · 2002Article
- Differential screening technology in the service of ovarian biology.Reviews in endocrine & metabolic disorders · 2002Review
- A critical evaluation of differential display as a tool to identify genes involved in legume nodulation: looking back and looking forward.Nucleic acids research · 2001Article
- Progress in the application of DNA microarrays.Environmental health perspectives · 2001Review
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Corrections and comments
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Authors and funding
7 authors at 2 institutions in 2 countries.
Funding
Abstract
The identification of differential gene expressionbetween cells is a frequent goal in modern biological research. Here we demonstrate the coupling of representational difference analysis (RDA) of cDNA with microarray analysis of the output for high throughput screening. Two primary Ewing's sarcoma tissue samples with different biological behavior in vivo were compared by RDA: one which was metastatic and progressed rapidly; the other localized and successfully treated. A modified RDA protocol that minimizes the necessary starting material was employed. After a reduced number of subtractive rounds, the output of RDA was shotgun cloned into a plasmid vector. Inserts from individual colonies from the subtracted library were amplified with vector-specific primers and arrayed at high density on glass slides. The arrays were then hybridized with differentially fluorescently labeled starting amplicons from the two tissues and fluorescent signals were measured at each DNA spot. We show that the relative amounts of fluorescent signal correlate well with the abundance of fragments in the RDA amplicon and in the starting mRNA. In our system, we analyzed 192 products and 173 (90%) were appropriately detected as being >2-fold differentially expressed. Fifty unique, differentially expressed clones were identified. Therefore, the use of RDA essentially provides an enriched library of differentially expressed genes, while analysis of this library with microarrays allows rapid and reproducible screening of thousands of DNA molecules simultaneously. The coupling of these two techniques in this system resulted in a large pool of differentially expressed genes.
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