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

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Author(s): Hidetoshi Arakawa

Journal: Current Chemical Genomics
ISSN 1875-3973

Volume: 6;
Issue: 1;
Start page: 1;
Date: 2012;
Original page

Keywords: ptamer | conformational differences | fluorescence detection | intercalating dyes | microchip electrophoresis | SELEX.

ABSTRACT
Microchip electrophoresis (ME) coupled with fluorescence detection was used to estimate the binding activityof aptamer in each systematic evolution of ligands by exponential enrichment (SELEX) round for a target molecule. Thisapproach is a non-radioisotopic, rapid and simple platform, and electrophoretic separation appears to be an effective tech-nique for aptamers of oligonucleotide molecules. We tried to obtain gonadotropin-specific RNA aptamer by the above ap-proach. As a result, the peaks of aptamers based on the conformational differences between them were separated and de-tected on the electropherograms. Moreover, the intensity of peak of unbound aptamer was decreased with progressionthrough the SELEX rounds, suggesting that RNA aptamer with high affinity was obtained by the proposed method.

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