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Label-free and sensitive detection of micrococcal nuclease activity using DNA-scaffolded silver nanoclusters as a fluorescence indicator

机译:使用DNA支架的银纳米簇作为荧光指示剂,无标记且灵敏地检测微球菌核酸酶活性

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A novel and label-free method for micrococcal nuclease (MNase) detection has been presented based on single-stranded DNA (ssDNA)-scaffolded fluorescent silver nanoclusters (AgNCs). The ssDNA was introduced as the substrate for MNase and also as the scaffold for the synthesis of the AgNCs. In the absence of MNase, the ssDNA was not digested. As a result, the fluorescent AgNCs were formed and exhibited strong fluorescence. In the presence of MNase, the DNA was digested, which prohibited the formation of the AgNCs due to the lack of the DNA scaffold, resulting in weak fluorescence. The fluorescence intensity exhibits a linear correlation to MNase concentration in the range of 0 U mLa?’1 to 2 ?— 10a?’4 U mLa?’1 with a detection limit of 8 ?— 10a?’6 U mLa?’1. Given its simplicity, easy operation, sensitivity and cost-effectiveness, this method can be extended to other nuclease assays.
机译:基于单链DNA(ssDNA)支架的荧光银纳米簇(AgNCs),提出了一种新颖且无标记的微球菌核酸酶(MNase)检测方法。引入ssDNA作为MNase的底物,也作为AgNC合成的支架。在缺少MNase的情况下,不消化ssDNA。结果,形成了荧光AgNC并显示出强荧光。在存在MNase的情况下,DNA被消化,由于缺少DNA支架而阻止了AgNC的形成,从而导致荧光弱。荧光强度与MNase浓度呈线性相关,范围为0 U mLa?'1至2?-10a?'4 U mLa?'1,检出限为8?-10a?'6 U mLa?'1 。鉴于其简单,易操作,灵敏性和成本效益,该方法可以扩展到其他核酸酶检测。

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