TITLE:
A Novel Label-Free Fluorescence Strategy Based on Dumbbell Probe for Sensitive Detection of DNA Ligase
AUTHORS:
Wenping Zhu, Xinlu Wang, Liyan Dai, Weijie Yang, Yanxia Li, Ruilan Liu
KEYWORDS:
Dumbbell Probe, DNA Ligase, SYBR Green I, DNA Ligation Reaction, Exonuclease
JOURNAL NAME:
Open Access Library Journal,
Vol.8 No.10,
October
9,
2021
ABSTRACT: 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.