Rapid isolation of cosmids from defined subregions by differential Alu-PCR hybridization on chromosome 22-specific library.
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.
Zucman
J
J
Laboratoire de Génétique des Tumeurs, Institut Curie, Paris, France.
Delattre
O
O
Desmaze
C
C
Azambuja
C
C
Rouleau
G
G
De Jong
P
P
Aurias
A
A
Thomas
G
G
D000818:Animals; D002874:Chromosome Mapping; D002892:Chromosomes, Human, Pair 22; D003360:Cosmids; D006224:Cricetinae; D003412:Cricetulus; D015723:Gene Library; D006801:Humans; D006822:Hybrid Cells; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid