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Development of RNA Aptamer and its Ligand Binding Assay on Microchip Electrophoresis

Microchip electrophoresis (ME) coupled with fluorescence detection was used to estimate the binding activity of aptamer in each systematic evolution of ligands by exponential enrichment (SELEX) round for a target molecule. This approach is a non-radioisotopic, rapid and simple platform, and electrop...

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Autores principales: Ohno, Ken-ichi, Nakata, Chikara, Sano, Yoshihiro, Nishikawa, Fumiko, Nishikawa, Satoshi, Arakawa, Hidetoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bentham Open 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3267091/
https://www.ncbi.nlm.nih.gov/pubmed/22291866
http://dx.doi.org/10.2174/1875397301206010001
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author Ohno, Ken-ichi
Nakata, Chikara
Sano, Yoshihiro
Nishikawa, Fumiko
Nishikawa, Satoshi
Arakawa, Hidetoshi
author_facet Ohno, Ken-ichi
Nakata, Chikara
Sano, Yoshihiro
Nishikawa, Fumiko
Nishikawa, Satoshi
Arakawa, Hidetoshi
author_sort Ohno, Ken-ichi
collection PubMed
description Microchip electrophoresis (ME) coupled with fluorescence detection was used to estimate the binding activity of aptamer in each systematic evolution of ligands by exponential enrichment (SELEX) round for a target molecule. This approach is a non-radioisotopic, rapid and simple platform, and electrophoretic separation appears to be an effective technique for aptamers of oligonucleotide molecules. We tried to obtain gonadotropin-specific RNA aptamer by the above approach. As a result, the peaks of aptamers based on the conformational differences between them were separated and detected on the electropherograms. Moreover, the intensity of peak of unbound aptamer was decreased with progression through the SELEX rounds, suggesting that RNA aptamer with high affinity was obtained by the proposed method.
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spelling pubmed-32670912012-01-30 Development of RNA Aptamer and its Ligand Binding Assay on Microchip Electrophoresis Ohno, Ken-ichi Nakata, Chikara Sano, Yoshihiro Nishikawa, Fumiko Nishikawa, Satoshi Arakawa, Hidetoshi Curr Chem Genomics Article Microchip electrophoresis (ME) coupled with fluorescence detection was used to estimate the binding activity of aptamer in each systematic evolution of ligands by exponential enrichment (SELEX) round for a target molecule. This approach is a non-radioisotopic, rapid and simple platform, and electrophoretic separation appears to be an effective technique for aptamers of oligonucleotide molecules. We tried to obtain gonadotropin-specific RNA aptamer by the above approach. As a result, the peaks of aptamers based on the conformational differences between them were separated and detected on the electropherograms. Moreover, the intensity of peak of unbound aptamer was decreased with progression through the SELEX rounds, suggesting that RNA aptamer with high affinity was obtained by the proposed method. Bentham Open 2012-01-24 /pmc/articles/PMC3267091/ /pubmed/22291866 http://dx.doi.org/10.2174/1875397301206010001 Text en © Ohno et al.; Licensee Bentham Open. http://creativecommons.org/licenses/by-nc/3.0/ This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.
spellingShingle Article
Ohno, Ken-ichi
Nakata, Chikara
Sano, Yoshihiro
Nishikawa, Fumiko
Nishikawa, Satoshi
Arakawa, Hidetoshi
Development of RNA Aptamer and its Ligand Binding Assay on Microchip Electrophoresis
title Development of RNA Aptamer and its Ligand Binding Assay on Microchip Electrophoresis
title_full Development of RNA Aptamer and its Ligand Binding Assay on Microchip Electrophoresis
title_fullStr Development of RNA Aptamer and its Ligand Binding Assay on Microchip Electrophoresis
title_full_unstemmed Development of RNA Aptamer and its Ligand Binding Assay on Microchip Electrophoresis
title_short Development of RNA Aptamer and its Ligand Binding Assay on Microchip Electrophoresis
title_sort development of rna aptamer and its ligand binding assay on microchip electrophoresis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3267091/
https://www.ncbi.nlm.nih.gov/pubmed/22291866
http://dx.doi.org/10.2174/1875397301206010001
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