Document Detail


Locked nucleic acid-based in situ detection of microRNAs in mouse tissue sections.
MedLine Citation:
PMID:  17571058     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Gregor Obernosterer; Javier Martinez; Mattias Alenius
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nature protocols     Volume:  2     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2007  
Date Detail:
Created Date:  2007-06-15     Completed Date:  2007-08-14     Revised Date:  2008-03-24    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1508-14     Citation Subset:  IM    
Affiliation:
Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Dr Bohr-Gasse 3, 1030 Vienna, Austria.
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MeSH Terms
Descriptor/Qualifier:
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:
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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