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High-throughput gene discovery in the rat
Journal article   Open access   Peer reviewed

High-throughput gene discovery in the rat

Katrina Fishler, Jack Gardiner, Lankai Guo, Brad Johnson, Catherine Keppel, Rikki Kreger, Mark Lebeck, Rudy Marcelino, Vladan Miljkovich, Mindee Perdue, …
Genome research, Vol.14(4), pp.733-741
04/2004
DOI: 10.1101/gr.1414204
PMCID: PMC383320
PMID: 15060017
url
https://doi.org/10.1101/gr.1414204View
Published (Version of record) Open Access

Abstract

The rat is an important animal model for human diseases and is widely used in physiology. In this article we present a new strategy for gene discovery based on the production of ESTs from serially subtracted and normalized cDNA libraries, and we describe its application for the development of a comprehensive nonredundant collection of rat ESTs. Our new strategy appears to yield substantially more EST clusters per ESTs sequenced than do previous approaches that did not use serial subtraction. However, multiple rounds of library subtraction resulted in high frequencies of otherwise rare internally primed cDNAs, defining the limits of this powerful approach. To date, we have generated >200,000 3' ESTs from >100 cDNA libraries representing a wide range of tissues and developmental stages of the laboratory rat. Most importantly, we have contributed to approximately 50,000 rat UniGene clusters. We have identified, arrayed, and derived 5' ESTs from >30,000 unique rat cDNA clones. Complete information, including radiation hybrid mapping data, is also maintained locally at http://genome.uiowa.edu/clcg.html. All of the sequences described in this article have been submitted to the dbEST division of the NCBI.
Software Gene Library Humans DNA, Complementary - genetics Genes - genetics Rats Male Sequence Analysis, DNA - statistics & numerical data Rats, Sprague-Dawley Polyadenylation - genetics Animals RNA Processing, Post-Transcriptional - genetics Female Mice Sequence Analysis, DNA - methods Expressed Sequence Tags

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