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Biotechniques. 2001 Mar;30(3):512-6, 518-20.

Generation of full-length cDNA libraries enriched for differentially expressed genes for functional genomics.

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Clontech Laboratories, Inc., 1020 East Meadow Circle, Palo Alto, CA 94303, USA.


Here, we describe the application of a RecA-based cloning technology to generate full-length cDNA libraries enriched for genes that are differentially expressed between tumor and normal tissue samples. First, we show that the RecA-based method can be used to enrich cDNA libraries for several target genes in a single reaction. Then, we demonstrate that this method can be extended to enrich a cDNA library for many full-length cDNA clones using fragments derived from a subtracted cDNA population. The results of these studies show that this RecA-mediated cloning technology can be used to convert subtracted cDNAs or a mixture of several cDNA fragments corresponding to differentially expressed genes into a full-length library in a single reaction. This procedure yields a population of expression-ready clones that can be used for further high-throughput functional screening.

[Indexed for MEDLINE]

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