Document Detail


Efficient nucleic acid detection by templated reductive quencher release.
MedLine Citation:
PMID:  19886694     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
RNA-templated fluorescence activation is a nucleic acid detection strategy that offers the possibility of direct visual detection of genetic information in living cells. Here we describe a new reaction strategy for fluorescence activation in which a phosphine on one DNA probe reduces an azide group in a linker on a second probe, resulting in linker cleavage and release of a fluorescence quenching group. These "Q-STAR" probes are shown to yield a strong fluorescence turn-on signal in approximately 20 min, with very low background and substantial amplification by turnover on the template. A green/red pair of such probes allowed the discrimination of two bacterial species by a single nucleotide difference in their 16S rRNA. The beneficial properties of the reductive quencher release design make these probes promising candidates for widespread application in the detection of nucleic acids in vitro and in cells.
Authors:
Raphael M Franzini; Eric T Kool
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  131     ISSN:  1520-5126     ISO Abbreviation:  J. Am. Chem. Soc.     Publication Date:  2009 Nov 
Date Detail:
Created Date:  2009-11-05     Completed Date:  2010-01-07     Revised Date:  2010-12-03    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  16021-3     Citation Subset:  IM    
Affiliation:
Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA.
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MeSH Terms
Descriptor/Qualifier:
Azides
Bacteria / isolation & purification
Cells
DNA Probes / chemistry*
Fluorescence
Molecular Probe Techniques*
Nucleic Acids / analysis*
Phosphines
RNA, Ribosomal, 16S / genetics
Grant Support
ID/Acronym/Agency:
GM068122/GM/NIGMS NIH HHS; R01 GM068122-05/GM/NIGMS NIH HHS; R01 GM068122-06/GM/NIGMS NIH HHS; R01 GM068122-08/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Azides; 0/DNA Probes; 0/Nucleic Acids; 0/Phosphines; 0/RNA, Ribosomal, 16S

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


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