| Locked nucleic acid based beacons for surface interaction studies and biosensor development. | |
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MedLine Citation:
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PMID: 19351140 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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DNA sensors and microarrays permit fast, simple, and real-time detection of nucleic acids through the design and use of increasingly sensitive, selective, and robust molecular probes. Specifically, molecular beacons (MBs) have been employed for this purpose; however, their potential in the development of solid-surface-based biosensors has not been fully realized. This is mainly a consequence of the beacon's poor stability because of the hairpin structure once immobilized onto a solid surface, commonly resulting in a low signal enhancement. Here, we report the design of a new MB that overcomes some of the limitations of MBs for surface immobilization. Essentially, this new design adds locked nucleic acid bases (LNAs) to the beacon structure, resulting in a LNA molecular beacon (LMB) with robust stability after surface immobilization. To test the efficacy of LMBs against that of regular molecular beacons (RMBs), the properties of selectivity, sensitivity, thermal stability, hybridization kinetics, and robustness for the detection of target sequences were compared and evaluated. A 25-fold enhancement was achieved for the LMB on surface with detection limits reaching the low nanomolar range. In addition, the LMB-based biosensor was shown to possess better stability, reproducibility, selectivity, and robustness when compared to the RMB. Therefore, as an alternative to conventional DNA and as a prospective tool for use in both DNA microarrays and biosensors, these results demonstrate the potential of the locked nucleic acid bases for nucleic acid design for surface immobilization. |
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Authors:
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Karen Martinez; M-Carmen Estevez; Yanrong Wu; Joseph A Phillips; Colin D Medley; Weihong Tan |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Analytical chemistry Volume: 81 ISSN: 1520-6882 ISO Abbreviation: Anal. Chem. Publication Date: 2009 May |
Date Detail:
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Created Date: 2009-04-30 Completed Date: 2009-06-25 Revised Date: 2011-09-26 |
Medline Journal Info:
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Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
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Languages: eng Pagination: 3448-54 Citation Subset: IM |
Affiliation:
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Center for Research at the Bio/Nano Interface, Department of Chemistry, University of Florida, Gainesville, Florida 32611, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Base Sequence Biosensing Techniques / methods* Biotechnology Cattle Cell Extracts Kinetics Nucleic Acid Hybridization Nucleic Acid Probes / chemical synthesis*, chemistry, genetics Oligonucleotide Array Sequence Analysis Oligonucleotides / chemistry* Sensitivity and Specificity Surface Properties Temperature |
| Grant Support | |
ID/Acronym/Agency:
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R01 GM066137-01/GM/NIGMS NIH HHS; R01 GM066137-02/GM/NIGMS NIH HHS; R01 GM066137-03/GM/NIGMS NIH HHS; R01 GM066137-04/GM/NIGMS NIH HHS; R01 GM066137-04S1/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Cell Extracts; 0/Nucleic Acid Probes; 0/Oligonucleotides; 0/locked nucleic acid |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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