Novel molecular beacon assays for DNA cleavage reactions

Jianwei Jeffrey Li; Charles Zehui Cao; Ronald Geyer; Weihong Tan

doi:10.1117/12.424592

16 April 2001 Novel molecular beacon assays for DNA cleavage reactions

Jianwei Jeffrey Li, Charles Zehui Cao, Ronald Geyer, Weihong Tan

Proceedings Volume 4264, Genomics and Proteomics Technologies; (2001) https://doi.org/10.1117/12.424592
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

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.

Citation Download Citation

Jianwei Jeffrey Li, Charles Zehui Cao, Ronald Geyer, and Weihong Tan "Novel molecular beacon assays for DNA cleavage reactions", Proc. SPIE 4264, Genomics and Proteomics Technologies, (16 April 2001); https://doi.org/10.1117/12.424592

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