Acta Phys. -Chim. Sin. ›› 2017, Vol. 33 ›› Issue (10): 2052-2057.doi: 10.3866/PKU.WHXB201750105

• ARTICLE • Previous Articles     Next Articles

Study on the Synthesis of DNA via Rolling Circle Amplification

Shu-Zhen LIU,Zhi-Qing ZHANG*(),Fang WANG,Ting ZHOU,Xiu-Feng WANG,Gou-Dong ZHANG,Ting-Ting LIU,Hong-Zhi ZHANG   

  • Received:2017-03-29 Published:2017-07-17
  • Contact: Zhi-Qing ZHANG E-mail:zhangzq@upc.edu.cn
  • Supported by:
    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

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.

Key words: Rolling circle amplification, Nucleic acid, dNTPs, Polymerase, Primer

MSC2000: 

  • O648