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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Bentham Open
2012
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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. |
format | Online Article Text |
id | pubmed-3267091 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Bentham Open |
record_format | MEDLINE/PubMed |
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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