title: Full-length cDNAs: more than just reaching the ends.
abstract: 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.
authors.affiliation: Department of Biochemistry, McGill Cancer Center, McGill University, Montreal, Quebec, Canada H3G 1Y6.
authors.forename: M
authors.initials: M
authors.lastname: Das
authors.forename: I
authors.initials: I
authors.lastname: Harvey
authors.forename: L L
authors.initials: LL
authors.lastname: Chu
authors.forename: M
authors.initials: M
authors.lastname: Sinha
authors.forename: J
authors.initials: J
authors.lastname: Pelletier
mesh_terms: D001331:Automation; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis / Q000302:isolation & purification; D015723:Gene Library; D005822:Genetic Vectors; D015894:Genome, Human; D006801:Humans; D016366:Open Reading Frames; D012333:RNA, Messenger / Q000302:isolation & purification; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry; D020506:Untranslated Regions
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D020506:Untranslated Regions; D012194:RNA-Directed DNA Polymerase