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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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Detalles Bibliográficos
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
Descripción
Sumario: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.