Abstract
In order to identify transcribed sequences from large genomic regions we developed and applied direct screening of genomic reference libraries. Briefly, reference libraries in lambda or cosmid vectors are screened with cDNA probes depleted of highly repeated sequence elements. The cDNA probes can be made from the same or related species. Further, by repeating the screenings with cDNA probes made from different tissues, the pattern of expression of sequences in each clone can be inferred. To derive sequence from positive genomic clones, DNA from these are Southern blotted and hybridized with the depleted cDNA probe. Small fragments identified in these blots are then sequenced and these sequences analyzed by GRAIL (1), a neural network program which detects coding exons from genomic sequence. We demonstrate the efficient isolation of expressed sequences from reference libraries of whole chromosomes, chromosomal regions and YAC clones, using cDNA probes from the same and related species.
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© 1994 Springer Science+Business Media New York
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Schwabe, W., Lawrence, B.J., Robb, A.S., Hopfinger, R.M., Hochgeschwender, U., Brennan, M.B. (1994). Direct cDNA Screening of Genomic Reference Libraries - A Rapid Method for the Identification of Transcribed Sequences in Large Genomic Regions. In: Hochgeschwender, U., Gardiner, K. (eds) Identification of Transcribed Sequences. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-2562-2_14
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DOI: https://doi.org/10.1007/978-1-4615-2562-2_14
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