| Rapid, single-step nucleic acid detection. | |
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MedLine Citation:
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PMID: 18563395 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A rapid detection method for nucleic acid based on bioluminescence resonance energy transfer (BRET) from the luminescence donor Renilla luciferase to an acceptor quantum dot upon oligonucleotide probe hybridization has been developed. Utilizing a competitive assay, we detected the target nucleic acid by correlating the BRET signal with the amount of target present in the sample. This method allows for the detection of as little as 4 pmol (20 nM) of nucleic acid in a single-step, homogeneous format both in vitro in a buffer matrix as well as in a cellular matrix. Using this method, one may perform nucleic acid detection in as little as 30 min, showing much improvement over time-consuming blotting methods and solid-phase methods which require multiple wash steps to remove unbound probe. This is the first report on the use of quantum dots as a BRET acceptor in the development of a nucleic acid hybridization assay. |
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Authors:
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Kyle A Cissell; Sean Campbell; Sapna K Deo |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, Non-P.H.S. Date: 2008-06-19 |
Journal Detail:
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Title: Analytical and bioanalytical chemistry Volume: 391 ISSN: 1618-2650 ISO Abbreviation: Anal Bioanal Chem Publication Date: 2008 Aug |
Date Detail:
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Created Date: 2008-07-30 Completed Date: 2008-09-26 Revised Date: 2009-01-29 |
Medline Journal Info:
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Nlm Unique ID: 101134327 Medline TA: Anal Bioanal Chem Country: Germany |
Other Details:
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Languages: eng Pagination: 2577-81 Citation Subset: IM |
Affiliation:
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Department of Chemistry and Chemical Biology, Indiana University Purdue University Indianapolis, 402 N. Blackford St., Room LD 326, Indianapolis, IN 46202, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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DNA
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analysis* Energy Transfer Escherichia coli / chemistry, genetics Imidazoles / chemistry Luciferases, Renilla / chemistry* Luminescent Agents / chemistry* Luminescent Measurements / methods* Nucleic Acid Hybridization / methods Oligonucleotide Probes / chemistry Pyrazines / chemistry Quantum Dots* |
| Chemical | |
Reg. No./Substance:
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0/Imidazoles; 0/Luminescent Agents; 0/Oligonucleotide Probes; 0/Pyrazines; 55779-48-1/coelenterazine; 9007-49-2/DNA; EC 1.13.12.5/Luciferases, Renilla |
| Comments/Corrections | |
Erratum In:
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Anal Bioanal Chem. 2008 Nov;392(5):1011 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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