功能核酸筛选过程中次级文库的有效制备

doi:10.13560/j.cnki.biotech.bull.1985.2022-0743

摘要/Abstract

摘要：

通过SELEX技术体外筛选功能核酸元件是其发挥各种功能的必要前提。SELEX程序中，制备高质量的单链次级文库是最基础、最关键的技术之一，很大程度上决定了筛选的成败。全面总结了功能核酸筛选中的单链生成策略，如基于扩增的方法、酶消化技术、基于链霉亲和素包被珠的链分离和基于迁移率变化分离获得次级文库等，详细阐述了各种方法的优缺点与关键细节，以期为进一步开发高效的功能核酸筛选方法奠定基础。

关键词: 功能核酸, 筛选, 适配体, PCR, 单链DNA生成

Abstract:

In vitro screening of functional nucleic acids（FNAs）by SELEX is an essential prerequisite for their various functions. In the SELEX process, the preparation of high-quality single-strand secondary library is one of the most basic and critical technologies, which largely determines the success or failure of screening. Here, we comprehensively summarize the generation strategies of single-strand DNA for FNAs selection, such as amplification-based methods, enzymatic digestion techniques, magnetic separation with streptavidin-coated beads, and secondary library regeneration strategies based on mobility shift. The advantages, disadvantages and key details of various methods are elaborated with aiming to lay a foundation for the further development of efficient FNAs screening methods.

Key words: functional nucleic acids, screening, aptamer, PCR, generation of ssDNA

李天顺, 李宸葳, 王佳, 朱龙佼, 许文涛. 功能核酸筛选过程中次级文库的有效制备[J]. 生物技术通报, 2023, 39(3): 116-122.

LI Tian-shun, LI Chen-wei, WANG Jia, ZHU Long-Jiao, XU Wen-tao. Efficient Generation of Secondary Libraries During Functional Nucleic Acids Screening[J]. Biotechnology Bulletin, 2023, 39(3): 116-122.

图/表 1

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