利用DNA荧光发光适体作为序列特异性核酸分析信号报告的分裂杂交探针。

IF 1.2 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES
AnnaMarie Knowles, Justine Monsalve, Yulia Gerasimova
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引用次数: 0

摘要

与用于瞬时核酸检测的传统杂交探针不同,基于dna的荧光发光适体(FLAPs)提供了低成本的荧光信号读出,而不需要用荧光团和/或猝灭剂标记核酸信号报告者,因此在生物分析分析中很有前景。相反,FLAP非共价结合染料配体,其在水溶液中表现出固有的低荧光,但在FLAP结合后变得高发射。本协议描述了一种利用迄今为止报道的最有效的DNA FLAP - DAP-10-42设计分裂点亮适体传感器(SLASs)的算法。当配备与感兴趣的核酸靶点互补的核酸序列时,SLAS是一种很有前途的核酸分析工具,允许对核酸靶点进行序列特异性检测,选择性低至一个核苷酸,从而能够分析单核苷酸取代(SNSs)。SLASs提供无标签荧光信号读出的优点,可以用传统的基于比色管的荧光分光光度计,便携式荧光计进行测量,或在手持光源激发时进行视觉观察。SLAS方法有利于生物传感在疾病诊断、环境监测和生物分子研究中的应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis.

DNA-based fluorescent light-up aptamers (FLAPs) are promising for bioanalytical assays because they provide a low-cost fluorescent signal readout without the need for labeling of nucleic acid signal reporters with fluorophores and/or quenchers, unlike conventional hybridization probes used for instantaneous nucleic acid detection. Instead, FLAPs non-covalently bind dye ligands , which exhibit intrinsically low fluorescence in aqueous solutions, but become highly emitting upon FLAP binding. This protocol describes an algorithm to design split light-up aptamer sensors (SLASs) utilizing DAP-10-42, the most efficient DNA FLAP reported thus far. When equipped with nucleic acid sequences complementary to a nucleic acid target of interest, SLAS is a promising tool for nucleic acid analysis allowing for the sequence-specific detection of nucleic acid targets with selectivity down to one nucleotide to enable analysis of single-nucleotide substitutions (SNSs). SLASs offer the advantage of a label-free fluorescence-based signal readout, which can be measured with a conventional cuvette-based fluorescent spectrophotometer, a portable fluorometer, or visually observed upon excitation with a handheld light source. The SLAS approach is beneficial for biosensing applications in disease diagnostics, environmental monitoring, and biomolecular research.

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来源期刊
Jove-Journal of Visualized Experiments
Jove-Journal of Visualized Experiments MULTIDISCIPLINARY SCIENCES-
CiteScore
2.10
自引率
0.00%
发文量
992
期刊介绍: JoVE, the Journal of Visualized Experiments, is the world''s first peer reviewed scientific video journal. Established in 2006, JoVE is devoted to publishing scientific research in a visual format to help researchers overcome two of the biggest challenges facing the scientific research community today; poor reproducibility and the time and labor intensive nature of learning new experimental techniques.
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