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Nucleic Acids Res ; 20(6): 1401-4, 1992 Mar 25.
Artículo en Inglés | MEDLINE | ID: mdl-1561096

RESUMEN

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.


Asunto(s)
Cromosomas Humanos Par 5 , Cósmidos , Secuencia de Bases , Southern Blotting , Clonación Molecular , ADN , Biblioteca de Genes , Humanos , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa
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