High-Throughput Plasmid cDNA Library Screening

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Libraries of cDNA clones are valuable resources foranalysing the expression, structure, and regulation of genes, as well asfor studying protein functions and interactions. Full-length cDNA clonesprovide information about intron and exon structures, splice junctionsand 5'- and 3'-untranslated regions (UTRs). Open reading frames (ORFs)derived from cDNA clones can be used to generate constructs allowingexpression of native proteins and N- or C-terminally tagged proteins.Thus, obtaining full-length cDNA clones and sequences for most or allgenes in an organism is critical for understanding genome functions.Expressed sequence tag (EST) sequencing samples cDNA libraries at random,which is most useful at the beginning of large-scale screening projects.However, ... continued below

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Wan, Kenneth H.; Yu, Charles; George, Reed A.; Carlson, JosephW.; Hoskins, Roger A.; Svirskas, Robert et al. May 24, 2006.

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Libraries of cDNA clones are valuable resources foranalysing the expression, structure, and regulation of genes, as well asfor studying protein functions and interactions. Full-length cDNA clonesprovide information about intron and exon structures, splice junctionsand 5'- and 3'-untranslated regions (UTRs). Open reading frames (ORFs)derived from cDNA clones can be used to generate constructs allowingexpression of native proteins and N- or C-terminally tagged proteins.Thus, obtaining full-length cDNA clones and sequences for most or allgenes in an organism is critical for understanding genome functions.Expressed sequence tag (EST) sequencing samples cDNA libraries at random,which is most useful at the beginning of large-scale screening projects.However, as projects progress towards completion, the probability ofidentifying unique cDNAs via EST sequencing diminishes, resulting in poorrecovery of rare transcripts. We describe an adapted, high-throughputprotocol intended for recovery of specific, full-length clones fromplasmid cDNA libraries in five days.

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  • Journal Name: Nature Protocols; Journal Volume: 0; Journal Issue: 0; Related Information: Journal Publication Date: 2006

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  • Report No.: LBNL--60261
  • Grant Number: DE-AC02-05CH11231
  • Grant Number: NIHHG002673
  • Office of Scientific & Technical Information Report Number: 923335
  • Archival Resource Key: ark:/67531/metadc895530

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Office of Scientific & Technical Information Technical Reports

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  • May 24, 2006

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  • Sept. 27, 2016, 1:39 a.m.

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  • Oct. 3, 2017, 2 p.m.

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Wan, Kenneth H.; Yu, Charles; George, Reed A.; Carlson, JosephW.; Hoskins, Roger A.; Svirskas, Robert et al. High-Throughput Plasmid cDNA Library Screening, article, May 24, 2006; Berkeley, California. (digital.library.unt.edu/ark:/67531/metadc895530/: accessed September 20, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.