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Methylene blue covalently attached to single stranded DNA as electroactive label for potential bioassays

机译:亚甲基蓝共价连接到单链DNA上作为潜在生物测定的电活性标记

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摘要

Methylene blue is an electroactive molecule that has been employed for the detection of the DNA hybridization event in electrochemical sensors. However, its use as a covalent label is very scarce and in most of the cases, non-covalent interactions (hydrophobic, electrostatic) are employed. Although it has advantages as simplicity and fewer number of procedure steps, the covalent attachment is less exploited in the development of these sensors. In this article, the electrochemical behavior of methylene blue attached to different DNA-strands is studied. Several lengths (15- and 30-mer) and different degree of DNA modification (MB-DNA, MB-DNA-MB and MB-DNA-SH) have been studied. The highest signals were obtained for longer strands with two MB molecules. In all the cases the signal is enhanced by CNT-nanostructuration of the electrode. Adsorption on these modified screen-printed electrodes allowed the amplification by employing an accumulation time. In this way, a sensitivity of −0.2864 μA μM and a limit of detection of 800 nM for a 120 s accumulation time were obtained.
机译:亚甲蓝是一种电活性分子,已用于检测电化学传感器中的DNA杂交事件。然而,其用作共价标记的用途非常稀少,并且在大多数情况下,采用了非共价相互作用(疏水,静电)。尽管共价连接具有简单易行,操作步骤少等优点,但在这些传感器的开发中却很少采用共价连接。在本文中,研究了附着在不同DNA链上的亚甲蓝的电化学行为。已经研究了几种长度(15-和30-mer)和不同程度的DNA修饰(MB-DNA,MB-DNA-MB和MB-DNA-SH)。对于具有两个MB分子的较长链,获得了最高信号。在所有情况下,电极的CNT纳米结构都会增强信号。这些修饰的丝网印刷电极上的吸附可通过利用累积时间来进行扩增。这样,获得的灵敏度为-0.2864μAμM,在120累积时间内的检测限为800 nM。

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