title: Real-time monitoring of double-stranded DNA cleavage using molecular beacons.
abstract: Traditional methods to assay enzymatic cleavage of DNA are discontinuous, time-consuming and laborious. Here, we report a new approach for real-time monitoring of double-stranded DNA cleavage by restriction endonuclease based on nucleic acid ligation using molecular beacon. Upon cleavage of DNA, the cleavage product can be ligated by DNA ligase, which results in a fluorescence enhancement of the molecular beacon. This method permits real-time monitoring of DNA cleavage and makes it easy to characterize the activity of restriction endonuclease and to study the cleavage reaction kinetics.
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Yuelu-Qu District, Changsha 410082, Hunan, PR China.
authors.forename: Changbei
authors.initials: C
authors.lastname: Ma
authors.forename: Zhiwen
authors.initials: Z
authors.lastname: Tang
authors.forename: Xiqin
authors.initials: X
authors.lastname: Huo
authors.forename: Xiaohai
authors.initials: X
authors.lastname: Yang
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Weihong
authors.initials: W
authors.lastname: Tan
mesh_terms: D004247:DNA / Q000378:metabolism*; D053837:DNA Cleavage; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000378:metabolism; D015342:DNA Probes; D004262:DNA Restriction Enzymes / Q000378:metabolism; D005453:Fluorescence; D007700:Kinetics; D008722:Methods; D015336:Molecular Probe Techniques
chemical_list: D015342:DNA Probes; D004247:DNA; D004262:DNA Restriction Enzymes; D011088:DNA Ligases; D000072481:DNA Ligase ATP