Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (3): 116-122.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0743

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Efficient Generation of Secondary Libraries During Functional Nucleic Acids Screening

LI Tian-shun1(), LI Chen-wei1, WANG Jia2, ZHU Long-Jiao2, XU Wen-tao1,2()   

  1. 1. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083
    2. Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, Beijing 100193
  • Received:2022-06-21 Online:2023-03-26 Published:2023-04-10

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