| Locked nucleic acid-based in situ detection of microRNAs in mouse tissue sections. | |
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MedLine Citation:
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PMID: 17571058 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Here we describe a method for sensitive and specific histological detection of microRNAs (miRNAs) by in situ hybridization. The protocol focuses on the use of locked nucleic acids (LNAs), which are bi-cyclic RNA analogs that allow a significant increase in the hybridization temperature and thereby an enhanced stringency for short probes as required for miRNA detection. The protocol is optimized for cryosections in order to study the spatial and temporal expression of miRNAs with high sensitivity and resolution. We detail how to construct probes, set up and conduct an LNA in situ hybridization experiment. In addition, we discuss alternative colorimetric strategies that can be used to effectively detect and visualize miRNAs including double staining with other markers. Setting up and conducting the in situ experiment is estimated to take approximately 1 week, assuming that all the component parts are readily available. |
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Authors:
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Gregor Obernosterer; Javier Martinez; Mattias Alenius |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Nature protocols Volume: 2 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2007 |
Date Detail:
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Created Date: 2007-06-15 Completed Date: 2007-08-14 Revised Date: 2008-03-24 |
Medline Journal Info:
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Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
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Languages: eng Pagination: 1508-14 Citation Subset: IM |
Affiliation:
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Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Dr Bohr-Gasse 3, 1030 Vienna, Austria. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Gene Expression Regulation In Situ Hybridization / methods* Mice MicroRNAs / analysis*, chemistry*, genetics, metabolism Molecular Probe Techniques* Oligonucleotides Oligonucleotides, Antisense / analysis*, chemistry* Sensitivity and Specificity |
| Chemical | |
Reg. No./Substance:
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0/MicroRNAs; 0/Oligonucleotides; 0/Oligonucleotides, Antisense; 0/locked nucleic acid |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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