Real-time monitoring of double-stranded DNA cleavage using molecular beacons.
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.
Ma
Changbei
C
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Yuelu-Qu District, Changsha 410082, Hunan, PR China.
Tang
Zhiwen
Z
Huo
Xiqin
X
Yang
Xiaohai
X
Li
Wei
W
Tan
Weihong
W
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
D015342:DNA Probes; D004247:DNA; D004262:DNA Restriction Enzymes; D011088:DNA Ligases; D000072481:DNA Ligase ATP