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Article: Rapid generation of region-specific genomic clones by chromosome midrodissection: Isolation of DNA from a region frequently deleted in malignant melanoma

TitleRapid generation of region-specific genomic clones by chromosome midrodissection: Isolation of DNA from a region frequently deleted in malignant melanoma
Authors
Issue Date1992
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ygeno
Citation
Genomics, 1992, v. 14 n. 3, p. 680-684 How to Cite?
AbstractMalignant melanoma is frequently characterized by the deletion of the long arm of chromosome 6 (usually encompassing 6q16-q21). In an effort to saturate this region with DNA markers, microdissection and molecular cloning of DNA from banded human metaphases have been performed. This work was facilitated by the recent development of a novel chromosome microdissection scheme that omits microchemical manipulation of DNA. Microdissection was targeted on band 6q21. Direct PCR amplification of dissected DNA was first used as a probe in chromosomal in situ hybridization of normal metaphases to confirm the specificity of material excised for cloning. A genomic library of 20,000 clones, which is highly enriched for sequences encompassing 6q21, was then constructed. Clones from this library have been mapped against a human-rodent somatic cell hybrid mapping panel that divides chromosome 6 into seven regions, confirming the localization of probes within the target region. Direct PCR amplification of DNA excised by microdissection greatly simplifies and facilitates this chromosome band-specific cloning strategy. The isolation of microclones from this region of chromosome 6 should assist in establishing a physical map of the melanoma deletion region.
Persistent Identifierhttp://hdl.handle.net/10722/150695
ISSN
2014 Impact Factor: 2.284
2014 SCImago Journal Rankings: 1.457
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorGuan, XYen_US
dc.contributor.authorMeltzer, PSen_US
dc.contributor.authorCao, Jen_US
dc.contributor.authorTrent, JMen_US
dc.date.accessioned2012-06-26T06:08:53Z-
dc.date.available2012-06-26T06:08:53Z-
dc.date.issued1992en_US
dc.identifier.citationGenomics, 1992, v. 14 n. 3, p. 680-684en_US
dc.identifier.issn0888-7543en_US
dc.identifier.urihttp://hdl.handle.net/10722/150695-
dc.description.abstractMalignant melanoma is frequently characterized by the deletion of the long arm of chromosome 6 (usually encompassing 6q16-q21). In an effort to saturate this region with DNA markers, microdissection and molecular cloning of DNA from banded human metaphases have been performed. This work was facilitated by the recent development of a novel chromosome microdissection scheme that omits microchemical manipulation of DNA. Microdissection was targeted on band 6q21. Direct PCR amplification of dissected DNA was first used as a probe in chromosomal in situ hybridization of normal metaphases to confirm the specificity of material excised for cloning. A genomic library of 20,000 clones, which is highly enriched for sequences encompassing 6q21, was then constructed. Clones from this library have been mapped against a human-rodent somatic cell hybrid mapping panel that divides chromosome 6 into seven regions, confirming the localization of probes within the target region. Direct PCR amplification of DNA excised by microdissection greatly simplifies and facilitates this chromosome band-specific cloning strategy. The isolation of microclones from this region of chromosome 6 should assist in establishing a physical map of the melanoma deletion region.en_US
dc.languageengen_US
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ygenoen_US
dc.relation.ispartofGenomicsen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshChromosome Deletionen_US
dc.subject.meshChromosomes, Human, Pair 6 - Ultrastructureen_US
dc.subject.meshCloning, Molecular - Methodsen_US
dc.subject.meshDna, Single-Strandeden_US
dc.subject.meshHumansen_US
dc.subject.meshIn Situ Hybridization, Fluorescenceen_US
dc.subject.meshMelanoma - Geneticsen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshPolymerase Chain Reactionen_US
dc.titleRapid generation of region-specific genomic clones by chromosome midrodissection: Isolation of DNA from a region frequently deleted in malignant melanomaen_US
dc.typeArticleen_US
dc.identifier.emailGuan, XY:xyguan@hkucc.hku.hken_US
dc.identifier.authorityGuan, XY=rp00454en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/S0888-7543(05)80168-5en_US
dc.identifier.pmid1427895-
dc.identifier.scopuseid_2-s2.0-0026539935en_US
dc.identifier.volume14en_US
dc.identifier.issue3en_US
dc.identifier.spage680en_US
dc.identifier.epage684en_US
dc.identifier.isiWOS:A1992JW35200018-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridGuan, XY=7201463221en_US
dc.identifier.scopusauthoridMeltzer, PS=7102464641en_US
dc.identifier.scopusauthoridCao, J=36983493000en_US
dc.identifier.scopusauthoridTrent, JM=7201692482en_US

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