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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier B.V.
2007
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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. |
format | Online Article Text |
id | pubmed-7127251 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | Elsevier B.V. |
record_format | MEDLINE/PubMed |
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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