School of Science, China University of Petroleum(East China), Qingdao 266580, Shandong Province, P. R. China
基金项目：
the Scientific Research Foundation for Returned Overseas Chinese Scholars(2014010615);National Natural Science Foundation of China(21603276);National Natural Science Foundation of China(21303267);Natural Science Foundation of Shandong Province, China(ZR2016BL14);Fundamental Research Funds for the Central Universities, China
School of Science, China University of Petroleum(East China), Qingdao 266580, Shandong Province, P. R. China
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.
