Abstract
Although advances in the development of positional cloning techniques have rapidly accelerated the pace of physical mapping and gene localization, the complete and efficient isolation of transcribed sequences from within large targeted genomic regions remains a significant challenge. Here, we describe two approaches used to isolate transcripts encoded within region Xp11.21. First, subcloned genomic fragments derived from regional YAC clones were used to identify evolutionarily conserved sequences (ECSs); ECSs were used to screen cDNA libraries and isolate regional transcripts. Second, YAC DNA was immobilized on a membrane and hybridized to PCR-amplified cDNA clone inserts; selectively retained inserts were subcloned to construct an enriched region-specific cDNA library. Both techniques were successfully used to isolate region-specific transcribed sequences. The advantages and limitations of each approach are discussed.
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Burright, E.N., Pasteris, N.G., Bialecki, M.D., Gorski, J.L. (1994). Use of cDNA Selection and Evolutionarily Conserved Sequences to Isolate Transcribed Sequences from Region Xp11.21. In: Hochgeschwender, U., Gardiner, K. (eds) Identification of Transcribed Sequences. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-2562-2_3
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DOI: https://doi.org/10.1007/978-1-4615-2562-2_3
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