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Electrochemical Assay for deoxyribonuclease I Activity

机译:脱氧核糖核酸酶I活性的电化学分析

摘要

A thiolated oligonucleotide having three ferrocenes was immobilized on a gold electrode through the sulfur–gold linkage. This electrode showed a current response based on the redox reaction of the ferrocene moieties and this response was decreased after treatment with deoxyribonuclease I (DNase I), suggesting the disappearance of the ferrocene moieties on the electrode by the DNase I digestion. A linear correlation between i0 and i, which are current peaks before and after DNase I treatment, respectively, was observed and this slope was decreased with increase in the amount of DNase I. No current decrease was observed in the presence of EDTA or RNase A instead of DNase I. These results suggested that the current decrease responded specifically to the amount of DNase I and this electrode could be used for an electrochemical DNase I assay. Under the optimum conditions of DNase I digestion at 37 °C for 30 min, a quantitative analysis could be achieved in the range of 10−4–10−2 units/μl of DNase I.
机译:具有三个二茂铁的硫醇化寡核苷酸通过硫金键固定在金电极上。该电极显示出基于二茂铁部分的氧化还原反应的电流响应,并且在用脱氧核糖核酸酶I(DNase I)处理后该响应降低了,表明通过DNase I消化电极上的二茂铁部分消失了。观察到i0和i之间的线性相关性,分别是DNase I处理之前和之后的电流峰值,并且该斜率随DNase I量的增加而减小。在EDTA或RNase A存在的情况下,未观察到电流减小这些结果表明电流的减少对DNase I的量有特殊的反应,并且该电极可用于电化学DNase I分析。在DNase I在37°C下消化30分钟的最佳条件下,可以进行定量分析,其范围为10−4–10−2-2个单位/μlDNaseI。

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