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  • Title: Isolation of region-specific cosmids from chromosome 5 by hybridization with microdissection clones.
    Author: Saltman DL, Dolganov GM, Pearce BS, Kuo SS, Callahan PJ, Cleary ML, Lovett M.
    Journal: Nucleic Acids Res; 1992 Mar 25; 20(6):1401-4. PubMed ID: 1561096.
    Abstract:
    A method is described for the isolation of chromosome region specific cosmids. The 5q35 region of the long arm of human chromosome 5 was microdissected, digested with MboI, ligated to oligonucleotide adaptors, amplified by the polymerase chain reaction and cloned into a plasmid vector. Inserts which did not contain highly repetitive sequences were used to screen a chromosome 5 cosmid library by direct hybridization. There were 33 positive cosmid clones identified with 4 microclones. Individual cosmid clones were biotinylated and used as probes for fluorescence in situ hybridization to metaphase chromosomes. Of the 33 cosmids that were mapped, 29 localized to q35 and 4 to q34, demonstrating the specificity of the microdissection library and the cosmids.
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