SELEX技术中制备单链DNA的方法  

Method for Preparing Single Stranded DNA in SELEX Technology

作　　者：徐婷钰 林陈水[1] XU Tingyu;LIN Chenshui(College of Pharmacy,Zhejiang University of Technology,Hangzhou 310014,China)

机构地区：[1]浙江工业大学药学院,浙江杭州310014

出　　处：《生物化工》2024年第2期238-242,共5页Biological Chemical Engineering

摘　　要：制备单链DNA(ssDNA)是许多常用实验中的重要内容,是通过指数富集的配体系统(SELEX)进行体外筛选核酸适配体的关键步骤之一。目前已报道多种方式可通过双链DNA(dsDNA)制备ssDNA,包括链霉亲和素包被磁珠分离、不对称PCR、不等大小引物PCR、酶消化、不对称PCR结合酶消化和不对称乳液PCR等,而如何快速高效地制备ssDNA是研究重点。本文综述了常用的几种通过聚合酶链式反应(PCR)制备ssDNA的方法,对其进行综合分析。根据产物的纯度、操作难易、实验成本等方面综合考虑,选择产物高纯度、操作简便且实验成本较低的ssDNA制备方法,为具有不同需求的应用场景提供参考依据。The preparation of single stranded DNA(ssDNA)is an important aspect of many commonly used experiments,and is one of the key steps for in vitro screening of nucleic acid aptamers through a systemic evolution of ligands by exponential enrichment(SELEX)ligand system.At present,various methods have been reported to prepare ssDNA through double stranded DNA(dsDNA),including streptavidin coated magnetic bead separation,asymmetric PCR,unequal size primer PCR,enzyme digestion,asymmetric PCR binding enzyme digestion and asymmetric emulsion PCR etc,and how to quickly and efficiently prepare ssDNA is a research focus.This article reviews several commonly used methods for preparing ssDNA through polymerase chain reaction(PCR)and provides a comprehensive analysis of them.Based on the comprehensive consideration of product purity,ease of operation,and experimental cost,a ssDNA preparation method with high product purity,simple operation,and low experimental cost is selected,which will provide reference for application scenarios with different requirements.

关 键 词：核酸适配体 单链DNA制备 聚合酶链式反应 

分 类 号：Q812[生物学—生物工程]