High-throughput plasmid cDNA library screening.

@article{citeulike:1854225, abstract = {Libraries of cDNA clones are valuable resources for analyzing the expression, structure and regulation of genes, and for studying protein functions and interactions. Full-length cDNA clones provide information about intron and exon structures, splice junctions, and 5' and 3' untranslated regions (UTRs). Open reading frames (ORFs) derived from cDNA clones can be used to generate constructs allowing the expression of both wild-type proteins and proteins tagged at their amino or carboxy terminus. Thus, obtaining full-length cDNA clones and sequences for most or all genes in an organism is essential for understanding genome functions. EST sequencing samples cDNA libraries at random, an approach that is most useful at the beginning of large-scale screening projects. As projects progress towards completion, however, the probability of identifying unique cDNAs by EST sequencing diminishes, resulting in poor recovery of rare transcripts. Here we describe an adapted, high-throughput protocol intended for the recovery of specific, full-length clones from plasmid cDNA libraries in 5 d.}, address = {Department of Genome Sciences, Earnest Orlando Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, California 94720, USA.}, author = {Wan, K. H. and Yu, C. and George, R. A. and Carlson, J. W. and Hoskins, R. A. and Svirskas, R. and Stapleton, M. and Celniker, S. E. }, citeulike-article-id = {1854225}, doi = {10.1038/nprot.2006.90}, issn = {1750-2799}, journal = {Nat Protoc}, number = {2}, pages = {624--632}, posted-at = {2007-11-02 03:42:34}, priority = {2}, title = {High-throughput plasmid cDNA library screening.}, url = {http://dx.doi.org/10.1038/nprot.2006.90}, volume = {1}, year = {2006} }

TY - JOUR ID - citeulike:1854225 L3 - citeulike-article-id:1854225 TI - High-throughput plasmid cDNA library screening. JF - Nat Protoc VL - 1 IS - 2 SP - 624 EP - 632 AD - Department of Genome Sciences, Earnest Orlando Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, California 94720, USA. SN - 1750-2799 N2 - Libraries of cDNA clones are valuable resources for analyzing the expression, structure and regulation of genes, and for studying protein functions and interactions. Full-length cDNA clones provide information about intron and exon structures, splice junctions, and 5' and 3' untranslated regions (UTRs). Open reading frames (ORFs) derived from cDNA clones can be used to generate constructs allowing the expression of both wild-type proteins and proteins tagged at their amino or carboxy terminus. Thus, obtaining full-length cDNA clones and sequences for most or all genes in an organism is essential for understanding genome functions. EST sequencing samples cDNA libraries at random, an approach that is most useful at the beginning of large-scale screening projects. As projects progress towards completion, however, the probability of identifying unique cDNAs by EST sequencing diminishes, resulting in poor recovery of rare transcripts. Here we describe an adapted, high-throughput protocol intended for the recovery of specific, full-length clones from plasmid cDNA libraries in 5 d. AU - Wan, KH AU - Yu, C AU - George, RA AU - Carlson, JW AU - Hoskins, RA AU - Svirskas, R AU - Stapleton, M AU - Celniker, SE PY - 2006/// UR - http://dx.doi.org/10.1038/nprot.2006.90 ER -