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Exonuclease III assisted and label-free detection of mercury ion based on toehold strand displacement amplification strategy

机译:核酸外切酶III基于脚趾链置换扩增策略辅助和无标记的汞离子检测

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A label-free, exonuclease III (Exo III) assisted and cost-effective Hg2+ fluorometric assay based on a strand displacement amplification strategy was developed with high sensitivity and selectivity. Thyminea€“thymine (Ta€“T) mismatches between the toehold domains in hairpin DNA and assistant DNA can serve as specific recognition elements for Hg2+ binding with the help of Ta€“Hg2+a€“T base pairs to initiate a toehold-mediated strand displacement reaction. Exo III provides a reduced background signal, and the method is label-free using an intercalator, Sybr green I (SG). The number of Ta€“T mismatches, the concentration of SG and Exo III, the incubation temperature and time were all optimized. Under the optimized condition, the limit of detection for Hg2+ can be as low as 9.42 pM with a linear range from 0.01 to 1 nM. Moreover, this proposed strategy shows excellent selectivity, and was successfully applied for the detection of Hg2+ in laboratory tap water and Yanjing lake water samples in our university.
机译:基于链置换扩增策略的无标记核酸外切酶III(Exo III)辅助且具有成本效益的Hg2 +荧光测定法具有很高的灵敏度和选择性。发夹状DNA和辅助DNA的脚趾域之间的胸腺嘧啶(胸腺嘧啶)(Ta)错配可以在TaHg2 +αT碱基对的帮助下作为Hg2 +结合的特异性识别元件,从而启动一个脚趾-介导的链置换反应。 Exo III提供了降低的背景信号,使用嵌入剂Sybr green I(SG),该方法无标签。 TaTT错配的数目,SG和Exo III的浓度,孵育温度和时间均得到优化。在优化条件下,Hg2 +的检出限可低至9.42 pM,线性范围为0.01至1 nM。此外,该方法具有很好的选择性,并已成功应用于我校实验室自来水和燕京湖水样中Hg2 +的检测。

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