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Chem. J. Chinese Universities ›› 2003, Vol. 24 ›› Issue (10): 1761.

• Articles • Previous Articles     Next Articles

Real-time Visualization of Ligation Reaction Catalyzed by E.coli DNA Ligase Based on Molecular Beacon

LIU Ling-Feng, TANG Zhi-Wen, WANG Ke-Min, TAN Wei-Hong, LI Jun, GUO Qiu-Ping, MENG Xiang-Xian, HUANG Shan-Sheng, LI Du   

  1. State Key Laboratory of Chemo/Biosensing and Chemometrics, Institute of Biological Technology, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China
  • Received:2003-04-15 Online:2003-10-24 Published:2003-10-24

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Abstract: Nucleic acids ligation is a crucial step in replication and repair of DNA and in genetic recombination.Researches on DNA ligation have been generally based on gel electrophoresis and autoradiography in the past, which include complex work and are time-consuming.In this paper, a novel approach was developed for real-time monitoring of DNA ligation process.By using this approach, DNA ligation process can be monitored in real-time providing abundant, successive, dynamic and real-time data, which is of great importance for the mechanistic studies and dynamics research of DNA ligation.Ulteriorly, a qualification assay method for the ligase is developed, with a linear range from 4.0×10-6to 2.0×10-4U/μLand a detection limit of 4.0×10-6U/μL.

Key words: E.coli DNA ligase, DNA ligation, Molecular beacon, Real-time monitoring

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