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 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.
Cite this paper
Zhu, W. , Wang, X. , Dai, L. , Yang, W. , Li, Y. and Liu, R. (2021). A Novel Label-Free Fluorescence Strategy Based on Dumbbell Probe for Sensitive Detection of DNA Ligase. Open Access Library Journal, 8, e7983. doi: http://dx.doi.org/10.4236/oalib.1107983.
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