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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
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TitleRapid generation of region-specific genomic clones by chromosome midrodissection: Isolation of DNA from a region frequently deleted in malignant melanoma
 
AuthorsGuan, XY1
Meltzer, PS1
Cao, J1
Trent, JM1
 
Issue Date1992
 
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ygeno
 
CitationGenomics, 1992, v. 14 n. 3, p. 680-684 [How to Cite?]
DOI: http://dx.doi.org/10.1016/S0888-7543(05)80168-5
 
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.
 
ISSN0888-7543
2013 Impact Factor: 2.793
 
DOIhttp://dx.doi.org/10.1016/S0888-7543(05)80168-5
 
ISI Accession Number IDWOS:A1992JW35200018
 
DC FieldValue
dc.contributor.authorGuan, XY
 
dc.contributor.authorMeltzer, PS
 
dc.contributor.authorCao, J
 
dc.contributor.authorTrent, JM
 
dc.date.accessioned2012-06-26T06:08:53Z
 
dc.date.available2012-06-26T06:08:53Z
 
dc.date.issued1992
 
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.
 
dc.description.naturelink_to_subscribed_fulltext
 
dc.identifier.citationGenomics, 1992, v. 14 n. 3, p. 680-684 [How to Cite?]
DOI: http://dx.doi.org/10.1016/S0888-7543(05)80168-5
 
dc.identifier.doihttp://dx.doi.org/10.1016/S0888-7543(05)80168-5
 
dc.identifier.epage684
 
dc.identifier.isiWOS:A1992JW35200018
 
dc.identifier.issn0888-7543
2013 Impact Factor: 2.793
 
dc.identifier.issue3
 
dc.identifier.pmid1427895
 
dc.identifier.scopuseid_2-s2.0-0026539935
 
dc.identifier.spage680
 
dc.identifier.urihttp://hdl.handle.net/10722/150695
 
dc.identifier.volume14
 
dc.languageeng
 
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ygeno
 
dc.publisher.placeUnited States
 
dc.relation.ispartofGenomics
 
dc.subject.meshBase Sequence
 
dc.subject.meshCells, Cultured
 
dc.subject.meshChromosome Deletion
 
dc.subject.meshChromosomes, Human, Pair 6 - Ultrastructure
 
dc.subject.meshCloning, Molecular - Methods
 
dc.subject.meshDna, Single-Stranded
 
dc.subject.meshHumans
 
dc.subject.meshIn Situ Hybridization, Fluorescence
 
dc.subject.meshMelanoma - Genetics
 
dc.subject.meshMolecular Sequence Data
 
dc.subject.meshPolymerase Chain Reaction
 
dc.titleRapid generation of region-specific genomic clones by chromosome midrodissection: Isolation of DNA from a region frequently deleted in malignant melanoma
 
dc.typeArticle
 
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<contributor.author>Meltzer, PS</contributor.author>
<contributor.author>Cao, J</contributor.author>
<contributor.author>Trent, JM</contributor.author>
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<date.available>2012-06-26T06:08:53Z</date.available>
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<description.abstract>Malignant 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.</description.abstract>
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<subject.mesh>Cloning, Molecular - Methods</subject.mesh>
<subject.mesh>Dna, Single-Stranded</subject.mesh>
<subject.mesh>Humans</subject.mesh>
<subject.mesh>In Situ Hybridization, Fluorescence</subject.mesh>
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Author Affiliations
  1. University of Michigan Medical School