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Nucleic Acids Research, 1992, Vol. 20, No. 6 1401-1404
© 1992

MOLECULAR BIOLOGY

Isolation of region-specific cosmids from chromosome 5 by hybridization with microdissection clones

David L. Saltman, Gregory M. Dolganov, Brian S. Pearce, Sofia S. Kuo1, Peter J. Callahan, Michael L. CIeary1 and Michael Lovett

Department of Molecular Genetics, Genelabs Incorporated 505 Penobscot Drive, Redwood City, CA 94063, USA 1Laboratory of Experimental Oncology, Department of Pathology, Stanford University Medical Center Stanford, CA 94305, USA

Received October 2, 1991. Revised February 17, 1992. Accepted February 17, 1992.

A method is described for the isolation of chromosome region specific cosmids. The 5q35 region of the long arm of human chromosome 5 was microdlssected, digested with Mbol, 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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