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dc.contributor.authorMelloni, Richard H.
dc.contributor.authorAronin, Neil
dc.contributor.authorDeGennaro, Louis J.
dc.contributor.authorFerris, Craig F.
dc.contributor.authorHarrison, Robert J.
dc.date2022-08-11T08:09:01.000
dc.date.accessioned2022-08-23T16:15:52Z
dc.date.available2022-08-23T16:15:52Z
dc.date.issued1997-05-01
dc.date.submitted2008-11-21
dc.identifier.citation<p>J Histochem Cytochem. 1997 May;45(5):755-63.</p>
dc.identifier.issn0022-1554 (Print)
dc.identifier.doi10.1177/002215549704500514
dc.identifier.pmid9154163
dc.identifier.urihttp://hdl.handle.net/20.500.14038/34194
dc.description.abstractWe present a novel procedure for detection of low- and high-abundance messenger RNAs in the brain by in situ hybridization histochemistry, by using fragmented double-stranded cDNA as molecular probes. The procedure involves digesting the cDNA of interest with the restriction endonuclease from Desulfocibrio desulfuricans (Dde I digestion), followed by random primed labeling, which generates a family of high specific activity cDNA fragments. This procedure is a rapid, straightforward, and reproducible method of obtaining sensitive probes for in situ hybridization and is generally applicable to the analysis of the expression of a large number of genes. Here we report the use of this procedure to prepare probes for the detection of synapsin I, p150Glued, neurotensin, c-fos, and c-jun mRNAs in brain, using both isotopic and non-isotopic labeling methods. Because this procedure does not require complex recombinant DNA manipulations or oligonucleotide design, it should prove useful to the non-molecular biologist examining the expression of genes in the central nervous system.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9154163&dopt=Abstract">Link to article in PubMed</a></p>
dc.relation.urlhttps://doi.org/10.1177/002215549704500514
dc.subjectAnimals; Brain; DNA Fragmentation; *DNA Probes; DNA, Complementary; Deoxyribonucleases, Type II Site-Specific; Hippocampus; *In Situ Hybridization; Microtubule-Associated Proteins; Neurotensin; Proto-Oncogene Proteins c-fos; Proto-Oncogene Proteins c-jun; RNA, Messenger; Rats; Rats, Sprague-Dawley; Synapsins
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleDde-I restriction endonuclease fragmentation: a novel method of generating cDNA probes for in situ hybridization in brain
dc.typeJournal Article
dc.source.journaltitleThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
dc.source.volume45
dc.source.issue5
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/gsbs_sp/851
dc.identifier.contextkey670507
html.description.abstract<p>We present a novel procedure for detection of low- and high-abundance messenger RNAs in the brain by in situ hybridization histochemistry, by using fragmented double-stranded cDNA as molecular probes. The procedure involves digesting the cDNA of interest with the restriction endonuclease from Desulfocibrio desulfuricans (Dde I digestion), followed by random primed labeling, which generates a family of high specific activity cDNA fragments. This procedure is a rapid, straightforward, and reproducible method of obtaining sensitive probes for in situ hybridization and is generally applicable to the analysis of the expression of a large number of genes. Here we report the use of this procedure to prepare probes for the detection of synapsin I, p150Glued, neurotensin, c-fos, and c-jun mRNAs in brain, using both isotopic and non-isotopic labeling methods. Because this procedure does not require complex recombinant DNA manipulations or oligonucleotide design, it should prove useful to the non-molecular biologist examining the expression of genes in the central nervous system.</p>
dc.identifier.submissionpathgsbs_sp/851
dc.contributor.departmentDepartment of Neurology and Cell Biology
dc.contributor.departmentGraduate School of Biomedical Sciences
dc.source.pages755-63


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