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Article: Generation of band-specific painting probes from a single microdissected chromosome

TitleGeneration of band-specific painting probes from a single microdissected chromosome
Authors
Issue Date1993
PublisherOxford University Press. The Journal's web site is located at http://hmg.oxfordjournals.org/
Citation
Human Molecular Genetics, 1993, v. 2 n. 8, p. 1117-1121 How to Cite?
AbstractWe have developed a modified strategy for the generation of regional probes for human chromosomes by microdissection and degenerate oligonucleotide primed PCR. This modification dramatically increases the efficiency of amplification by pretreatment of the dissected chromatin with topoisomerase I (Topo I) before PCR. This protocol has enabled us to construct region-specific probes for fluorescence in situ hybridization (FISH) from a single microdissected chromosome. Results are presented which convincingly demonstrate that this new method generates high intensity region-specific FISH probes, while at the same time significantly decreasing the time-consuming and labor-intensive aspects of microdissection. The reduction of the number of copies required to generate a useful probe also significantly decreases the risk of contamination during the microdissection process. We believe this advance will allow microdissection to be more widely used in the cytogenetic analysis of chromosome rearrangements in both cancer and hereditary diseases. In addition, this method now makes it possible to construct a series of non-overlapping band-specific DNA microclone libraries to provide complete coverage of individual chromosomes for physical mapping.
Persistent Identifierhttp://hdl.handle.net/10722/150701
ISSN
2015 Impact Factor: 5.985
2015 SCImago Journal Rankings: 4.288
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorGuan, XYen_US
dc.contributor.authorTrent, JMen_US
dc.contributor.authorMeltzer, PSen_US
dc.date.accessioned2012-06-26T06:08:59Z-
dc.date.available2012-06-26T06:08:59Z-
dc.date.issued1993en_US
dc.identifier.citationHuman Molecular Genetics, 1993, v. 2 n. 8, p. 1117-1121en_US
dc.identifier.issn0964-6906en_US
dc.identifier.urihttp://hdl.handle.net/10722/150701-
dc.description.abstractWe have developed a modified strategy for the generation of regional probes for human chromosomes by microdissection and degenerate oligonucleotide primed PCR. This modification dramatically increases the efficiency of amplification by pretreatment of the dissected chromatin with topoisomerase I (Topo I) before PCR. This protocol has enabled us to construct region-specific probes for fluorescence in situ hybridization (FISH) from a single microdissected chromosome. Results are presented which convincingly demonstrate that this new method generates high intensity region-specific FISH probes, while at the same time significantly decreasing the time-consuming and labor-intensive aspects of microdissection. The reduction of the number of copies required to generate a useful probe also significantly decreases the risk of contamination during the microdissection process. We believe this advance will allow microdissection to be more widely used in the cytogenetic analysis of chromosome rearrangements in both cancer and hereditary diseases. In addition, this method now makes it possible to construct a series of non-overlapping band-specific DNA microclone libraries to provide complete coverage of individual chromosomes for physical mapping.en_US
dc.languageengen_US
dc.publisherOxford University Press. The Journal's web site is located at http://hmg.oxfordjournals.org/en_US
dc.relation.ispartofHuman Molecular Geneticsen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshChromatin - Ultrastructureen_US
dc.subject.meshChromosome Bandingen_US
dc.subject.meshChromosomes, Human - Ultrastructureen_US
dc.subject.meshDna Primersen_US
dc.subject.meshDna Probesen_US
dc.subject.meshDna Topoisomerases, Type Ien_US
dc.subject.meshHumansen_US
dc.subject.meshIn Situ Hybridization, Fluorescenceen_US
dc.subject.meshKaryotypingen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshPolymerase Chain Reaction - Methodsen_US
dc.titleGeneration of band-specific painting probes from a single microdissected chromosomeen_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.pmid8401492-
dc.identifier.scopuseid_2-s2.0-0027279527en_US
dc.identifier.volume2en_US
dc.identifier.issue8en_US
dc.identifier.spage1117en_US
dc.identifier.epage1121en_US
dc.identifier.isiWOS:A1993LV83600004-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridGuan, XY=7201463221en_US
dc.identifier.scopusauthoridTrent, JM=7201692482en_US
dc.identifier.scopusauthoridMeltzer, PS=7102464641en_US

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