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  • Title: Rapid isolation of cosmids from defined subregions by differential Alu-PCR hybridization on chromosome 22-specific library.
    Author: Zucman J, Delattre O, Desmaze C, Azambuja C, Rouleau G, De Jong P, Aurias A, Thomas G.
    Journal: Genomics; 1992 Jun; 13(2):395-401. PubMed ID: 1612598.
    Abstract:
    A method based on the differential screening of a chromosome-specific cosmid library with amplified inter-Alu sequences obtained from a set of somatic cell hybrids has been developed to target the isolation of probes from predefined subchromosomal regions. As a model system, we have used a chromosome 22-specific cosmid library and four cell hybrids containing different parts of this chromosome. The procedure has identified cosmids that demonstrate differential hybridization signals with Alu-PCR products from these cell hybrids. We show, by in situ hybridization or individual mapping, that their hybridization pattern is indicative of their sublocalization on chromosome 22, thus resulting in a large enrichment factor for the isolation of probes from specific small chromosome subregions. Depending on the local Alu-sequence density, from 3 to 10 independent loci per megabase of genome can thus be identified.
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