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Oligonucleotide Functionalised Microbeads: Indispensable Tools for High-Throughput Aptamer Selection

The functionalisation of microbeads with oligonucleotides has become an indispensable technique for high-throughput aptamer selection in SELEX protocols. In addition to simplifying the separation of binding and non-binding aptamer candidates, microbeads have facilitated the integration of other tech...

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Autores principales: Fraser, Lewis A., Kinghorn, Andrew B., Tang, Marco S. L., Cheung, Yee-Wai, Lim, Bryce, Liang, Shaolin, Dirkzwager, Roderick M., Tanner, Julian A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6332362/
https://www.ncbi.nlm.nih.gov/pubmed/26633328
http://dx.doi.org/10.3390/molecules201219766
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author Fraser, Lewis A.
Kinghorn, Andrew B.
Tang, Marco S. L.
Cheung, Yee-Wai
Lim, Bryce
Liang, Shaolin
Dirkzwager, Roderick M.
Tanner, Julian A.
author_facet Fraser, Lewis A.
Kinghorn, Andrew B.
Tang, Marco S. L.
Cheung, Yee-Wai
Lim, Bryce
Liang, Shaolin
Dirkzwager, Roderick M.
Tanner, Julian A.
author_sort Fraser, Lewis A.
collection PubMed
description The functionalisation of microbeads with oligonucleotides has become an indispensable technique for high-throughput aptamer selection in SELEX protocols. In addition to simplifying the separation of binding and non-binding aptamer candidates, microbeads have facilitated the integration of other technologies such as emulsion PCR (ePCR) and Fluorescence Activated Cell Sorting (FACS) to high-throughput selection techniques. Within these systems, monoclonal aptamer microbeads can be individually generated and assayed to assess aptamer candidate fitness thereby helping eliminate stochastic effects which are common to classical SELEX techniques. Such techniques have given rise to aptamers with 1000 times greater binding affinities when compared to traditional SELEX. Another emerging technique is Fluorescence Activated Droplet Sorting (FADS) whereby selection does not rely on binding capture allowing evolution of a greater diversity of aptamer properties such as fluorescence or enzymatic activity. Within this review we explore examples and applications of oligonucleotide functionalised microbeads in aptamer selection and reflect upon new opportunities arising for aptamer science.
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spelling pubmed-63323622019-01-24 Oligonucleotide Functionalised Microbeads: Indispensable Tools for High-Throughput Aptamer Selection Fraser, Lewis A. Kinghorn, Andrew B. Tang, Marco S. L. Cheung, Yee-Wai Lim, Bryce Liang, Shaolin Dirkzwager, Roderick M. Tanner, Julian A. Molecules Review The functionalisation of microbeads with oligonucleotides has become an indispensable technique for high-throughput aptamer selection in SELEX protocols. In addition to simplifying the separation of binding and non-binding aptamer candidates, microbeads have facilitated the integration of other technologies such as emulsion PCR (ePCR) and Fluorescence Activated Cell Sorting (FACS) to high-throughput selection techniques. Within these systems, monoclonal aptamer microbeads can be individually generated and assayed to assess aptamer candidate fitness thereby helping eliminate stochastic effects which are common to classical SELEX techniques. Such techniques have given rise to aptamers with 1000 times greater binding affinities when compared to traditional SELEX. Another emerging technique is Fluorescence Activated Droplet Sorting (FADS) whereby selection does not rely on binding capture allowing evolution of a greater diversity of aptamer properties such as fluorescence or enzymatic activity. Within this review we explore examples and applications of oligonucleotide functionalised microbeads in aptamer selection and reflect upon new opportunities arising for aptamer science. MDPI 2015-12-01 /pmc/articles/PMC6332362/ /pubmed/26633328 http://dx.doi.org/10.3390/molecules201219766 Text en © 2015 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Review
Fraser, Lewis A.
Kinghorn, Andrew B.
Tang, Marco S. L.
Cheung, Yee-Wai
Lim, Bryce
Liang, Shaolin
Dirkzwager, Roderick M.
Tanner, Julian A.
Oligonucleotide Functionalised Microbeads: Indispensable Tools for High-Throughput Aptamer Selection
title Oligonucleotide Functionalised Microbeads: Indispensable Tools for High-Throughput Aptamer Selection
title_full Oligonucleotide Functionalised Microbeads: Indispensable Tools for High-Throughput Aptamer Selection
title_fullStr Oligonucleotide Functionalised Microbeads: Indispensable Tools for High-Throughput Aptamer Selection
title_full_unstemmed Oligonucleotide Functionalised Microbeads: Indispensable Tools for High-Throughput Aptamer Selection
title_short Oligonucleotide Functionalised Microbeads: Indispensable Tools for High-Throughput Aptamer Selection
title_sort oligonucleotide functionalised microbeads: indispensable tools for high-throughput aptamer selection
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6332362/
https://www.ncbi.nlm.nih.gov/pubmed/26633328
http://dx.doi.org/10.3390/molecules201219766
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