Label-free Fluorescence Assay for Real-Time Monitoring of Site-Specific DNA Cleavage by EcoRI Endonu

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In this paper, we demonstrated a fluorescent assay for lable-free, facile, and real-time detection DNA cleavage by EcoRI endonuclease using SYBR Green I (SGI) as signal probe. The fluorescence of SGI dramatically increased when the free SGI mixed with double-strands (dsDNA) substrate. While EcoRI was added into the mixture of dsDNA and SGI, the fluorescence of SGI decreased because of DNA cleavage by EcoRI. EcoRI–DNA interaction was monitored real-time based on the alteration of fluorescence signal before and after digestion with EcoRI. Compared with previously reported methods, this approach is selective, simple, convenient and cost-efficient without any labelling of the probe or of the target.
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