DNA immobilization chemical interference due to aggregates study by Dip and Drop approach

Journal of biochemical and biophysical methods Pub Date : 2007-08-01 DOI:10.1016/j.jbbm.2007.05.006

Ram Ajore , Rakesh Kumar , Inderpreet Kaur , R.C. Sobti , Lalit M. Bharadwaj

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引用次数: 3

Abstract

In the present manuscript, we report the studies and observations for chemical interference due to aggregates formation during covalent immobilization of thiolated λ-DNA between gold microelectrodes. Dip and Drop approaches were employed to study DNA immobilization using thiolated oligos (oligoA 5′ GGGCGGCGACCT 3′ and oligoB 5′ AGGTCGCCGCCC 3′). As a result of aggregation, less interference was observed in Dip approach as compared to Drop approach. Atomic Force Microscopy (AFM) analysis of piranha treated gold surface revealed 47.5% increase in height roughness, contributing in interference by creating active sites. Cyclic voltammetry (CV) studies ascertain the multitude of adsorption states existing in long strand of DNA on surface. Surface coverage was found to be ∼ 72% (1.35 × 10¹⁰ molecules/cm²), and ∼ 42% (7.89 × 10⁹ molecules/ cm²) in Dip and Drop approach, respectively. Dip approach can be used as a measure to minimize interference due to aggregation.

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聚集体对DNA固定的化学干扰研究

在本文中，我们报告了在金微电极之间的硫代化λ-DNA共价固定过程中由于聚集物形成的化学干扰的研究和观察。采用浸渍法和滴注法研究硫代寡核苷酸(oligoA 5’GGGCGGCGACCT 3’和oligoB 5’AGGTCGCCGCCC 3’)的DNA固定。由于聚集的结果，Dip方法比Drop方法观察到的干扰少。原子力显微镜(AFM)分析了水虎鱼处理过的金表面，发现高度粗糙度增加了47.5%，这是由于产生了活性位点而造成的干扰。循环伏安法(CV)研究确定了DNA长链表面存在多种吸附态。在Dip和Drop方法中，表面覆盖率分别为~ 72% (1.35 × 1010分子/cm2)和~ 42% (7.89 × 109分子/cm2)。倾角法可以用来作为一种措施，以尽量减少由于聚集的干扰。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Journal of biochemical and biophysical methods

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