基于核酸外切酶(Exo I)选择性降解单链DNA的性质,为增加DNA折纸结构纯化方法的多样性,开发了一种快速除去剩余DNA单链、纯化DNA折纸结构的新方法。以三角形和矩形DNA折纸结构为研究对象,对缓冲液环境、酶用量、反应温度、反应时间等实验条件进行了优化,采用琼脂糖凝胶电泳方法与原子力显微技术对产物进行分析与表征。结果表明,该方法可以达到快速除去DNA单链、纯化DNA折纸结构的目的,Exo I酶处理不影响DNA折纸结构的完整性及后续的应用开发。

In order to increase the diversity of purification methods for DNA origami structure,a new method for fast purification of DNA origami nanostructures is developed by taking advantage of the property of selectively digestion of single-stranded DNA by Exonuclease I(Exo I).Triangle and rectangle-shaped DNA origami nanostructures are used for testing the hypothesis.The experimental conditions,including the usage of exonuclease I,buffer environment,reaction time and temperature,are optimized.Agarose gel electrophoresis and atomic force microscopy (AFM) techniques are employed to characterize DNA origami samples.The gel results and AFM data demonstrate that the Exo I-based method can be utilized for fast removing excess staple oligonucleotides and thus purifying the DNA origami nanostructures.

雷云翔(1996-),男,硕士研究生,研究方向为生物化工、DNA纳米技术,2019110488@mail.hfut.edu.cn。