title: Label-free molecular beacon for real-time monitoring of DNA polymerase activity.
abstract: Traditional methods for assaying DNA polymerase activity are discontinuous, time consuming, and laborious. Here, we report a new approach for label-free and real-time monitoring of DNA polymerase activity using a Thioflavin T (ThT) probe. In the presence of DNA polymerase, the DNA primer could be elongated through polymerase reaction to open MB1, leading to the release of the G-quartets. These then bind to ThT to form ThT/G-quadruplexes with an obvious fluorescence generation. It exhibits a satisfying detection result for the activity of DNA polymerase with a low detection limit of 0.05 unit/ml. In addition, no labeling with a fluorophore or a fluorophore-quencher pair is required; this method is fairly simple, fast, and low cost. Furthermore, the proposed method was also applied to assay the inhibition of DNA polymerase activity. This approach may offer potential applications in drug screening, clinical diagnostics, and some other related biomedical research.
authors.affiliation: State Key Laboratory of Medical Genetics and School of Life Sciences, Central South University, Changsha, 410013, China. macb2012@csu.edu.cn.
authors.forename: Changbei
authors.initials: C
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Medical Genetics and School of Life Sciences, Central South University, Changsha, 410013, China.
authors.forename: Haisheng
authors.initials: H
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Medical Genetics and School of Life Sciences, Central South University, Changsha, 410013, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Medical Genetics and School of Life Sciences, Central South University, Changsha, 410013, China.
authors.forename: Shunxin
authors.initials: S
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha, 410081, China.
authors.forename: Kemin
authors.initials: K
authors.lastname: Wang
mesh_terms: D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D006801:Humans
chemical_list: D004259:DNA-Directed DNA Polymerase
keywords: DNA polymerase assay; Label free; Molecular beacon; Real-time monitoring; Thioflavin T