Hefei National Laboratory for Physical Science at the Microscale, Core Facility Center for Life Sciences, School of Life Sciences, University of Science and Technology of China, Hefei 230026, P. R. China.
Analyst. 2017 Aug 21;142(17):3136-3139. doi: 10.1039/c7an01131h.
The development of a simple, low-cost, time-saving and universally applicable method to monitor the progression of aptamer selection is particularly challenging. Herein, a combined strategy dependent on quantitative polymerase chain reaction amplification curve (AC) and melting curve analysis (MCA) is developed to monitor the convergence of the aptamer species during selection progress. As a parallel and complementary method to affinity tests and binding analyses, the AC-MCA method can be used to achieve the DNA complexity assay when affinity is undetectable. It is independent of the target properties and SELEX methods. Therefore, it has been demonstrated as a universal monitoring tool in different SELEX methods towards different targets (small molecules, proteins, bacteria and cancer cells). The AC-MCA method will facilitate current new aptamer discovery and aptamer-based wide application.
开发一种简单、低成本、省时且通用的方法来监测适体选择的进展特别具有挑战性。在此,提出了一种依赖于定量聚合酶链反应扩增曲线 (AC) 和熔解曲线分析 (MCA) 的组合策略,以监测选择过程中适体种类的收敛。作为亲和性测试和结合分析的平行和互补方法,当亲和性不可检测时,AC-MCA 方法可用于实现 DNA 复杂度测定。它独立于目标性质和 SELEX 方法。因此,它已被证明是针对不同目标(小分子、蛋白质、细菌和癌细胞)的不同 SELEX 方法中的通用监测工具。AC-MCA 方法将促进当前新适体的发现和基于适体的广泛应用。