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A Novel Label-Free Fluorescence Strategy Based on Dumbbell Probe for Sensitive Detection of DNA Ligase

Wenping Zhu, Xinlu Wang, Liyan Dai, Weijie Yang, Yanxia Li, Ruilan Liu
2021 OALib  

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Based on the principle that DNA ligation reaction mediates hydrolysis of the dumbbell probe (DP), a novel label-free strategy for sensitive detection of DNA ligase was developed. DNA ligase ligated the nick on DP to generate a fully closed dumbbell DNA structure, which could prevent the hydrolysis of exonuclease, after combining with SYBR Green I (SG I), a strong fluorescent signal was obtained. In the absence of DNA ligase, DP was hydrolyzed into single nucleotides by exonuclease, resulting in
more » ... a rather weak signal. The linear range of this method for detecting T4 DNA ligase is 0.00004 -0.004 U/μL, and the detection limit is 0.00003 U/μL. The proposed strategy is sensitive, inexpensive and easy to operate, which may offer an effective tool for further applications in new drug screening.
doi:10.4236/oalib.1107983 fatcat:ytpsesx6yffl7nktwjikrh3ah4
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Wenping Zhu, Xinlu Wang, Liyan Dai, Weijie Yang, Yanxia Li, Ruilan Liu. "A Novel Label-Free Fluorescence Strategy Based on Dumbbell Probe for Sensitive Detection of DNA Ligase." OALib 08.10 (2021) 1-9