| A rational approach in probe design for nucleic acid-based biosensing. | |
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MedLine Citation:
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PMID: 21719269 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Development of nucleic acid-based sensing attracts the interest of many researchers in the field of both basic and applied research in chemistry. Major factors for the fabrication of a successful nucleic acid sensor include the design of probes for target sequence hybridization and their immobilization on the chip surface. Here we demonstrate that a rational choice of bioprobes has important impact on the sensor's analytical performances. Computational evaluations, by a simple and freely available program, successfully led to the design of the best probes for a given target, with direct application to nucleic acid-based sensing. We developed here an optimized and reproducible strategy for in silico probe design supported by optical transduction experiments. In particular Surface Plasmon Resonance imaging (SPRi), at the forefront of optical sensing, was used here as proof of principle. Five probes were selected, immobilized on gold chip surfaces by widely consolidated thiol chemistry and tested to validate the computational model. Using SPRi as the transducting component, real-time and label free analysis was performed, taking the Homo sapiens actin beta (ACTB) gene fragment as model system in nucleic acid detection. The experimental sensor behavior was further studied by evaluating the strength of the secondary structure of probes using melting experiments. Dedicated software was also used to evaluate probes' folding, to support our criteria. The SPRi experimental results fully validate the computational evaluations, revealing this approach highly promising as a useful tool to design biosensor probes with optimized performances. |
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Authors:
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M L Ermini; S Scarano; R Bini; M Banchelli; D Berti; M Mascini; M Minunni |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-6-12 |
Journal Detail:
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Title: Biosensors & bioelectronics Volume: - ISSN: 1873-4235 ISO Abbreviation: - Publication Date: 2011 Jun |
Date Detail:
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Created Date: 2011-7-1 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9001289 Medline TA: Biosens Bioelectron Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2011 Elsevier B.V. All rights reserved. |
Affiliation:
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Dipartimento di Chimica "Ugo Schiff", Università degli Studi di Firenze, Via della Lastruccia 13, 50019, Sesto F.no (FI), Italy. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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