物理化学学报2017，Vol.33Issue(10)：2052-2057,6.DOI:10.3866/PKU.WHXB201750105
DNA的滚环扩增合成研究
Study on the Synthesis of DNA via Rolling Circle Amplification
摘要
Abstract
Rolling circle amplification (RCA) is a simple and efficient isothermal enzymatic reaction,which has developed as a novel technology in the field of nucleic acid amplification.Its product has a wide range of applications in the assembly and preparation of multi-functional materials.Here,we report the effects of reaction time and the concentrations of deoxyribonucleoside triphosphates (dNTPs),polymerase,and primer on the product of RCA.The RCA product was characterized by methods including agarose gel electrophoresis,ultraviolet spectroscopy,and transmission electron microscopy (TEM).The results showed that the length of the RCA product was significantly affected by the reaction time,especially when the reaction time was less than 30 min.With an increase of dNTPs concentrations,the concentration and chain length of the RCA product increased.However,while the concentrations of enzyme and primer had little effect on the length of the RCA product,they had a large effect on its concentration.It is worth noting that the content of the RCA product decreased significantly in the presence of excess enzyme concentration.关键词
滚环扩增技术/核酸/dNTPs/聚合酶/引物Key words
Rolling circle amplification/Nucleic acid/dNTPs/Polymerase/Primer分类
化学化工引用本文复制引用
刘淑贞,张志庆,王芳,周亭,王秀凤,张国栋,刘婷婷,张洪芝..DNA的滚环扩增合成研究[J].物理化学学报,2017,33(10):2052-2057,6.基金项目
The project was supported by the Scientific Research Foundation for Returned Overseas Chinese Scholars (2014010615),National Natural Science Foundation of China (21603276,21303267),Natural Science Foundation of Shandong Province,China (ZR2016BL14),and Fundamental Research Funds for the Central Universities,China 教育部留学回国人员科研启动基金(2014010615),国家自然科学青年基金(21603276,21303267),山东省自然科学基金(ZR2016BL14),中央高校基本科研业务费资助项目 （2014010615）
