文章基本信息

标题：Quantitative selection of DNA aptamers through microfluidic selection and high-throughput sequencing
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作者：Minseon Cho ; Yi Xiao ; Jeff Nie 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2010
卷号：107
期号：35
页码：15373-15378
DOI：10.1073/pnas.1009331107
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：We describe the integration of microfluidic selection with high-throughput DNA sequencing technology for rapid and efficient discovery of nucleic acid aptamers. The Quantitative Selection of Aptamers through Sequencing method tracks the copy number and enrichment-fold of more than 10 million individual sequences through multiple selection rounds, enabling the identification of high-affinity aptamers without the need for the pool to fully converge to a small number of sequences. Importantly, this method allows the discrimination of sequences that arise from experimental biases rather than true high-affinity target binding. As a demonstration, we have identified aptamers that specifically bind to PDGF-BB protein with Kd < 3 nM within 3 rounds. Furthermore, we show that the aptamers identified by Quantitative Selection of Aptamers through Sequencing have [~]3-8-fold higher affinity and [~]2-4-fold higher specificity relative to those discovered through conventional cloning methods. Given that many biocombinatorial libraries are encoded with nucleic acids, we extrapolate that our method may be extended to other types of libraries for a range of molecular functions.
关键词：high-throughput DNA sequencing ; PDGF-BB ; Quantitative Selection of Aptamers through Sequencing ; MicroMagnetic Separation device