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Citation: LIU Shu-Zhen, ZHANG Zhi-Qing, WANG Fang, ZHOU Ting, WANG Xiu-Feng, ZHANG Gou-Dong, LIU Ting-Ting, ZHANG Hong-Zhi. Study on the Synthesis of DNA via Rolling Circle Amplification[J]. Acta Physico-Chimica Sinica, ;2017, 33(10): 2052-2057. doi: 10.3866/PKU.WHXB201750105 shu

Study on the Synthesis of DNA via Rolling Circle Amplification

School of Science, China University of Petroleum(East China), Qingdao 266580, Shandong Province, P. R. China

Received Date: 29 March 2017
Revised Date: 2 May 2017

Fund Project: 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.

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.
- Rolling circle amplification,
- Nucleic acid,
- dNTPs,
- Polymerase,
- Primer
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