Isolation of a cosmid sublibrary for a region of chromosome 12 frequently amplified in human cancers using a complex chromosome microdissection probe

Journal Article · Thu Feb 01 04:00:00 EST 1996 · Genomics

DOI:https://doi.org/10.1006/geno.1996.0057· OSTI ID:508334

Elkahloun, A G; Meltzer, P S; Guan, Xin-Yuan ^[1]

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1. National Institutes of Health, Bethesda, MD (United States); and others

Chromosome-specific cosmid libraries are in extremely useful resource for positional cloning projects. Once a particular region of interest has been identified, it would be of value to have an approach for isolating chromosome band-specific cosmids that could be assembled into a sublibrary for rapid screening. We constructed a region-specific sublibrary of 700 cosmids by screening a chromosome 12-specific cosmid library with a complex probe generated by degenerate oligonucleotide-primed PCR of a microdissected homogeneously staining region containing sequences amplified from chromosome 12q13-q15. Based on fluorescence in situ hybridization, approximately 60% of the cosmids in the sublibrary were derived from the microdissected region. To demonstrate further the utility of the sublibrary, a 150-kb contig containing the SAS and CDK4 genes was constructed, as well as several additional contigs between CDK4 and MDM2. This study demonstrates the possibility of utilizing probes generated by microdissection for assembling band-specific libraries that are amendable to rapid screening with multiple markers.

• OSTI ID: 508334
• Journal Information: Genomics, Journal Name: Genomics Journal Issue: 3 Vol. 31; ISSN 0888-7543; ISSN GNMCEP
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
99 GENERAL AND MISCELLANEOUS
BIOLOGICAL MARKERS
COMPILED DATA
COSMIDS
DESIGN
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
FLUORESCENCE
GENE AMPLIFICATION
GENES
GENETIC MAPPING
GROWTH
HUMAN CHROMOSOME 12
NEOPLASMS
OLIGONUCLEOTIDES
POLYMERASE CHAIN REACTION
PROBES
SARCOMAS
SCREENING