Document Detail


Modifications of RNase protection assay for neuroscience applications.
MedLine Citation:
PMID:  9821645     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This report describes how certain modifications of the Ribonuclease (RNase) protection assay may increase its efficiency by decreasing the time and cost of the procedure without compromising reliability. We show that, under the experimental conditions tested, the RNA samples can be precipitated by a solution of Tri Reagent in ethanol immediately following the RNase digestion step. Drying the samples under vacuum before dissolving them in the gel loading buffer improves the consistency of the assay as compared to air drying. Although these modifications are applicable to the RNase protection assay in general, we present an example that used a multiprobe set we developed and have used effectively in the analysis of cytokine mRNA regulation in the brain.
Authors:
S E Ilyin; D Gayle; C R Plata-Salamán
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of neuroscience methods     Volume:  84     ISSN:  0165-0270     ISO Abbreviation:  J. Neurosci. Methods     Publication Date:  1998 Oct 
Date Detail:
Created Date:  1999-01-26     Completed Date:  1999-01-26     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7905558     Medline TA:  J Neurosci Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  139-41     Citation Subset:  IM    
Affiliation:
Division of Molecular Biology, School of Life and Health Sciences, University of Delaware, Newark 19716-2590, USA. 78742@udel.eud
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MeSH Terms
Descriptor/Qualifier:
Animals
Brain / metabolism
Cytokines / genetics
Gene Expression Regulation
Indicators and Reagents
Liver / metabolism
Neurosciences / methods*
RNA / analysis*,  isolation & purification
RNA, Messenger / analysis,  genetics
Rats
Reproducibility of Results
Ribonucleases / metabolism*
Spleen / metabolism
Chemical
Reg. No./Substance:
0/Cytokines; 0/Indicators and Reagents; 0/RNA, Messenger; 63231-63-0/RNA; EC 3.1.-/Ribonucleases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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