工业微生物2025，Vol.55Issue(1)：110-115,124,7.DOI:10.3969/j.issn.1001-6678.2025.01.031
尿素变性PAGE中ssDNA回收方法的研究与优化
Study and Optimization of ssDNA Recovery Method in Urea Denaturation PAGE
摘要
Abstract
Obtaining high-purity and high-recovery ssDNA from denaturing PAGE is a crucial step in functional nucleic acid research and aptamer screening.In this article,four methods were employed to extract and recover ssDNA from urea denaturation PAGE,including direct extraction method,improved boiling method,kit recovery,and n-butanol recovery.The results demonstrated that the improved boiling method exhibited significantly higher recovery efficiency compared to the other three methods.After further optimization of experimental conditions,the recovery efficiency of ssDNA reached 70.33±0.35％,with increased purity(A260/A280=1.85).It indicated that this method can offer advantages such as simplicity in operation,rapid recovery speed,absence of interference from organic reagents,low cost(less than 1 yuan per recovery),and hold great potentials for practical applications in functional nucleic acid screening and recycling.关键词
尿素变性PAGE/ssDNA回收/改良煮沸法Key words
urea denaturation PAGE/ssDNA recovery/improved boiling method引用本文复制引用
郑楚君,黄海轩,王雨萱,叶煜玮,徐斐,袁敏..尿素变性PAGE中ssDNA回收方法的研究与优化[J].工业微生物,2025,55(1):110-115,124,7.基金项目
国家重点研发计划"食品营养与安全关键技术研发"重点专项(编号:2022YFF1101102). （编号:2022YFF1101102）
