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Proceedings Paper

Novel molecular beacon assays for DNA cleavage reactions
Author(s): Jianwei Jeffrey Li; Charles Zehui Cao; Ronald Geyer; Weihong Tan
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Paper Abstract

DNA cleavage reactions are important in both cellular processes and biotechnology. Conventional assays are discontinuous, time consuming and laborious, and there is a great demand for continuous, convenient and sensitive assays for these reactions. Here we introduce a novel method for the monitoring of the cleavage reactions in real time. Molecular beacon is used as a substrate for the single strand specific DNA nucleases. The cleavage of the molecular beacon by Si nuclease is demonstrated as an example for this method. The cleavage causes a great increase in fluorescence intensity of the molecular beacon, giving a signal-to-noise ratio of about 40. This assay is sensitive, continuous and easy to use. It can be applied to the detection of DNA nucleases, the characterization of these enzymes and for detailed study of the mechanisms of the cleavage reactions. The influence of solution conditions, such as macromolecular crowding, on the cleavage reactions has been studied.

Paper Details

Date Published: 16 April 2001
PDF: 8 pages
Proc. SPIE 4264, Genomics and Proteomics Technologies, (16 April 2001); doi: 10.1117/12.424592
Show Author Affiliations
Jianwei Jeffrey Li, Univ. of Florida (United States)
Charles Zehui Cao, Univ. of Florida (United States)
Ronald Geyer, Univ. of Florida (United States)
Weihong Tan, Univ. of Florida (United States)

Published in SPIE Proceedings Vol. 4264:
Genomics and Proteomics Technologies
Ramesh Raghavachari; Weihong Tan, Editor(s)

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