功能核酸筛选过程中次级文库的有效制备  

Efficient Generation of Secondary Libraries During Functional Nucleic Acids Screening

作　　者：李天顺 李宸葳 王佳[2] 朱龙佼 许文涛[1,2] LI Tian-shun;LI Chen-wei;WANG Jia;ZHU Long-Jiao;XU Wen-tao(College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083;Key Laboratory of Precision Nutrition and Food Quality,Department of Nutrition and Health,China Agricultural University,Beijing 100193)

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083 [2]中国农业大学营养与健康系,食品精准营养与质量控制教育部重点实验室,北京100193

出　　处：《生物技术通报》2023年第3期116-122,共7页Biotechnology Bulletin

基　　金：河北省重点研发计划项目(21372801D);青年人才托举工程项目(BYESS2022133);北京市景观休闲农业创新团队(BAIC09-2022)。

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。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.

关 键 词：功能核酸 筛选 适配体 PCR 单链DNA生成 

分 类 号：Q78[生物学—分子生物学]