| miRNA in situ hybridization in formaldehyde and EDC-fixed tissues. | |
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MedLine Citation:
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PMID: 19137005 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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MicroRNAs are small regulatory RNAs with many biological functions and disease associations. We showed that in situ hybridization (ISH) using conventional formaldehyde fixation results in substantial microRNA loss from mouse tissue sections, which can be prevented by fixation with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide that irreversibly immobilizes the microRNA at its 5' phosphate. We determined optimal hybridization parameters for 130 locked nucleic acid probes by recording nucleic acid melting temperature during ISH. |
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Authors:
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John T G Pena; Cherin Sohn-Lee; Sara H Rouhanifard; Janos Ludwig; Markus Hafner; Aleksandra Mihailovic; Cindy Lim; Daniel Holoch; Philipp Berninger; Mihaela Zavolan; Thomas Tuschl |
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Publication Detail:
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Type: Comparative Study; Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Date: 2009-01-11 |
Journal Detail:
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Title: Nature methods Volume: 6 ISSN: 1548-7105 ISO Abbreviation: Nat. Methods Publication Date: 2009 Feb |
Date Detail:
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Created Date: 2009-01-30 Completed Date: 2009-02-23 Revised Date: 2011-09-26 |
Medline Journal Info:
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Nlm Unique ID: 101215604 Medline TA: Nat Methods Country: United States |
Other Details:
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Languages: eng Pagination: 139-41 Citation Subset: IM |
Affiliation:
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Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, 1230 York Avenue, Box 186, New York, New York 10065, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Brain / cytology, metabolism Brain Chemistry Carbodiimides / chemistry* Formaldehyde / chemistry* In Situ Hybridization / methods* Mice MicroRNAs / analysis*, genetics Microscopy, Fluorescence / methods Neurons / cytology, metabolism Tissue Fixation / methods* |
| Grant Support | |
ID/Acronym/Agency:
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EY18082-01A2/EY/NEI NIH HHS; GM073047/GM/NIGMS NIH HHS; MH080442/MH/NIMH NIH HHS; P01 GM073047-04/GM/NIGMS NIH HHS; P01 GM073047-040001/GM/NIGMS NIH HHS; P01 GM073047-05/GM/NIGMS NIH HHS; P01 GM073047-050001/GM/NIGMS NIH HHS; R01 MH080442-01/MH/NIMH NIH HHS; R01 MH080442-02/MH/NIMH NIH HHS; R01 MH080442-03/MH/NIMH NIH HHS; //Howard Hughes Medical Institute; //Howard Hughes Medical Institute |
| Chemical | |
Reg. No./Substance:
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0/1-ethyl-3-(3-(diethylamino)propyl)carbodiimide; 0/Carbodiimides; 0/MicroRNAs; 50-00-0/Formaldehyde |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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