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DNA single-base mismatch study with an electrochemical enzymatic genosensor

A thorough selectivity study of DNA hybridization employing an electrochemical enzymatic genosensor is discussed here. After immobilizing on a gold film a 30-mer 3′-thiolated DNA strand, hybridization with a biotinylated complementary one takes place. Then, alkaline phosphatase is incorporated to th...

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Autores principales: Abad-Valle, Patricia, Fernández-Abedul, M.Teresa, Costa-García, Agustín
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7127251/
https://www.ncbi.nlm.nih.gov/pubmed/16950611
http://dx.doi.org/10.1016/j.bios.2006.07.015
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author Abad-Valle, Patricia
Fernández-Abedul, M.Teresa
Costa-García, Agustín
author_facet Abad-Valle, Patricia
Fernández-Abedul, M.Teresa
Costa-García, Agustín
author_sort Abad-Valle, Patricia
collection PubMed
description A thorough selectivity study of DNA hybridization employing an electrochemical enzymatic genosensor is discussed here. After immobilizing on a gold film a 30-mer 3′-thiolated DNA strand, hybridization with a biotinylated complementary one takes place. Then, alkaline phosphatase is incorporated to the duplex through the interaction streptavidin–biotin. Enzymatic generation of indigo blue from 3-indoxyl phosphate and subsequent electrochemical detection was made. The influence of hybridization conditions was studied in order to better discern between fully complementary and mismatched strands. Detection of 3, 2 and 1 mismatch was possible. The type and location of the single-base mismatch, as well as the influence of the length of the strands was studied too. Mutations that suppose displacement of the reading frame were also considered. The effect of the concentration on the selectivity was tested, resulting a highly selective genosensor with an adequate sensitivity and stability.
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spelling pubmed-71272512020-04-08 DNA single-base mismatch study with an electrochemical enzymatic genosensor Abad-Valle, Patricia Fernández-Abedul, M.Teresa Costa-García, Agustín Biosens Bioelectron Article A thorough selectivity study of DNA hybridization employing an electrochemical enzymatic genosensor is discussed here. After immobilizing on a gold film a 30-mer 3′-thiolated DNA strand, hybridization with a biotinylated complementary one takes place. Then, alkaline phosphatase is incorporated to the duplex through the interaction streptavidin–biotin. Enzymatic generation of indigo blue from 3-indoxyl phosphate and subsequent electrochemical detection was made. The influence of hybridization conditions was studied in order to better discern between fully complementary and mismatched strands. Detection of 3, 2 and 1 mismatch was possible. The type and location of the single-base mismatch, as well as the influence of the length of the strands was studied too. Mutations that suppose displacement of the reading frame were also considered. The effect of the concentration on the selectivity was tested, resulting a highly selective genosensor with an adequate sensitivity and stability. Elsevier B.V. 2007-03-15 2006-09-01 /pmc/articles/PMC7127251/ /pubmed/16950611 http://dx.doi.org/10.1016/j.bios.2006.07.015 Text en Copyright © 2006 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Abad-Valle, Patricia
Fernández-Abedul, M.Teresa
Costa-García, Agustín
DNA single-base mismatch study with an electrochemical enzymatic genosensor
title DNA single-base mismatch study with an electrochemical enzymatic genosensor
title_full DNA single-base mismatch study with an electrochemical enzymatic genosensor
title_fullStr DNA single-base mismatch study with an electrochemical enzymatic genosensor
title_full_unstemmed DNA single-base mismatch study with an electrochemical enzymatic genosensor
title_short DNA single-base mismatch study with an electrochemical enzymatic genosensor
title_sort dna single-base mismatch study with an electrochemical enzymatic genosensor
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7127251/
https://www.ncbi.nlm.nih.gov/pubmed/16950611
http://dx.doi.org/10.1016/j.bios.2006.07.015
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