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Physiol. Genomics 6: 57-80, 2001;
1094-8341/01 $5.00
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Received 1 May 2001; accepted in final form 23 May 2001.
Physiological Genomics 6:57-80 (2001)
1094-8341/01 $5.00 © 2001 American Physiological Society

Review

Full-length cDNAs: more than just reaching the ends

MANJULA DAS 1, ISABELLE HARVEY 1, LEE LEE CHU 1, MANISHA SINHA 1 and JERRY PELLETIER 1,2

1 Department of Biochemistry
2 McGill Cancer Center, McGill University, Montreal, Quebec, Canada H3G 1Y6

The development of functional genomic resources is essential to understand and utilize information generated from genome sequencing projects. Central to the development of this technology is the creation of high-quality cDNA resources and improved technologies for analyzing coding and noncoding mRNA sequences. The isolation and mapping of cDNAs is an entrée to characterizing the information that is of significant biological relevance in the genome of an organism. However, a bottleneck is often encountered when attempting to bring to full-length (or at least full-coding) a number of incomplete cDNAs in parallel, since this involves the nonsystematic, time consuming, and labor-intensive iterative screening of a number of cDNA libraries of variable quality and/or directed strategies to process individual clones (e.g., 5' rapid amplification of cDNA ends). Here, we review the current state of the art in cDNA library generation, as well as present an analysis of the different steps involved in cDNA library generation.

gene hunting; complementary DNA cloning; expressed sequence tags; expression validated gene




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