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

机译:RNA适体的开发及其在芯片电泳中的配体结合测定

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摘要

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.
机译:微芯片电泳(ME)与荧光检测相结合,用于通过目标分子的指数富集(SELEX)轮估计配体在每个系统进化中的适体结合活性。该方法是非放射性同位素,快速且简单的平台,电泳分离似乎是寡核苷酸分子适体的有效技术。我们试图通过上述方法获得促性腺激素特异性RNA适体。结果,基于它们之间的构象差异的适体的峰被分离并在电泳图上检测到。此外,未结合的适体的峰强度随着通过SELEX回合的进行而降低,这表明通过所提出的方法获得了具有高亲和力的RNA适体。

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