适体
指数富集配体系统进化
化学
荧光
流式细胞术
数字聚合酶链反应
单元格排序
分子生物学
聚合酶链反应
生物化学
核糖核酸
生物
基因
体外
物理
量子力学
作者
Xiaona Fang,Wenjing Li,Tian Gao,Qurat ul ain Zahra,Luo Z,Renjun Pei
出处
期刊:Talanta
[Elsevier]
日期:2022-05-01
卷期号:242: 123302-123302
被引量:8
标识
DOI:10.1016/j.talanta.2022.123302
摘要
In this paper, we report the development of a new strategy termed integrated digital PCR-fluorescence activated sorting based SELEX (IFS-SELEX) that enables rapid screening of aptamers against fluorescent targets. Initially, this strategy employs an integrated digital PCR system to amplify each sequence of a preliminarily enriched library, which is obtained by a traditional SELEX method, on the surface of polystyrene beads. Then, the as-prepared beads are incubated with the fluorescent target and then subjected to fluorescence-activated sorting. Since only those sequences with high binding affinity for the target are collected and sequenced, unnecessary analysis of ineligible sequences is avoided by this method, and the aptamer selection process is thereby greatly streamlined. As a proof-of-concept, we applied this strategy for the screening of aptamers against two fluorescent targets, i.e., ciprofloxacin (CFX) and thioflavin T (ThT), and successfully obtained corresponding sequences with low dissociation constants. The binding affinities of aptamers for ThT were well associated with the sorting regions defined in the fluorescence channel of the flow cytometry process. The experimental results demonstrated that the as-designed IFS-SELEX method can serve as a universal platform for rapid, facile, and efficient aptamer selection against various fluorescent targets.
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