Preparation of Quantum-Dot-Labeled DNA Substrate Materials
3:38
Microfluidic Device Construction
5:58
Surface Tethering of Quantum-Dot-Labeled DNA Substrate
7:07
REase-Mediated DNA Cleavage
7:51
Results: Single-Molecule TIRF Experiment for Parallelized Observations of DNA Cleavage Events
9:32
Conclusion
Transcript
С помощью этого протокола мы можем непосредственно наблюдать расщепление сайт-специфической ДНК эндонуклеазами рестрикции на уровне одной молекулы. Наш мультиплексный подход позволяет нам измерять время, необходимое сотням отдельных REases для зав
Sign in or start your free trial to access this content
Using quantum-dot-labeled DNA and total internal reflection fluorescence microscopy, we can investigate the reaction mechanism of restriction endonucleases while using unlabeled protein. This single-molecule technique allows for massively multiplexed observation of individual protein-DNA interactions, and data can be pooled to generate well-populated dwell-time distributions.