| Dde-I restriction endonuclease fragmentation: a novel method of generating cDNA probes for in situ hybridization in brain. | |
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MedLine Citation:
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PMID: 9154163 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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R H Melloni; N Aronin; L J DeGennaro; C F Ferris; R J Harrison |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society Volume: 45 ISSN: 0022-1554 ISO Abbreviation: J. Histochem. Cytochem. Publication Date: 1997 May |
Date Detail:
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Created Date: 1997-06-19 Completed Date: 1997-06-19 Revised Date: 2006-05-13 |
Medline Journal Info:
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Nlm Unique ID: 9815334 Medline TA: J Histochem Cytochem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 755-63 Citation Subset: IM |
Affiliation:
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Molecular Neurobiology Laboratory, University of Massachusetts Medical Center, Worcester 01655, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Brain / metabolism* DNA Fragmentation DNA Probes* DNA, Complementary Deoxyribonucleases, Type II Site-Specific / metabolism* Hippocampus / metabolism In Situ Hybridization* Microtubule-Associated Proteins / genetics, metabolism Neurotensin / genetics, metabolism Proto-Oncogene Proteins c-fos / genetics, metabolism Proto-Oncogene Proteins c-jun / genetics, metabolism RNA, Messenger Rats Rats, Sprague-Dawley Synapsins / genetics, metabolism |
| Chemical | |
Reg. No./Substance:
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0/DNA Probes; 0/DNA, Complementary; 0/Microtubule-Associated Proteins; 0/Proto-Oncogene Proteins c-fos; 0/Proto-Oncogene Proteins c-jun; 0/RNA, Messenger; 0/Synapsins; 144198-36-7/dynactin; 39379-15-2/Neurotensin; EC 3.1.21.-/endodeoxyribonuclease DdeI; EC 3.1.21.4/Deoxyribonucleases, Type II Site-Specific |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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