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Monitoring DNA Immobilization and Hybridization on Surfaces by Atomic Force Microscopy Force Measurements

机译:通过原子力显微镜力测量监测表面上的DNA固定和杂交

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DNA immobilization and hybridization was carried out on Au substrates that were modified with mercaptopropanoic acid and then treated with aluminum(III) solution. The positively charged Al(III) film can be used to immobilize both ds-DNA and ss-DNA. Atomic force microscopy (AFM) was used to monitor the process by force measurements between a negatively charged silica tip and the substrates while immersed in dilute electrolyte. Surface hybridization of ss-DNA produces an increase in the surface charge and surface potential of the substrates, which is reflected by the increasing repulsive force as determined from AFM force-separation curves. A single-base mismatch was detectable in surface hybridization. The AFM force measuring technique was also employed to investigate the interaction of (Ru(phen)_(3))~(2+) with ss-DNA and ds-DNA. The force measurement results showed that there is a small interaction between (Ru(phen)_(3))~(2+) and ss-DNA, which was ascribed to the electrostatic binding of (Ru(phen)_(3))~(2+) to the ss-DNA surface. For ds-DNA, there is a strong interaction which is believed to be due to the association or intercalation of (Ru(phen)_(3))~(2+) with ds-DNA.
机译:在用巯基丙酸修饰的Au底物上进行DNA固定和杂交,然后用铝(III)溶液处理。带正电的Al(III)膜可用于固定ds-DNA和ss-DNA。原子力显微镜(AFM)用于在浸入稀电解质的同时,通过带负电的硅胶头和基材之间的力测量来监控过程。 ss-DNA的表面杂交使底物的表面电荷和表面电势增加,这反映在由AFM力分离曲线确定的增加的排斥力上。在表面杂交中可检测到单碱基错配。原子力显微镜力测量技术还被用来研究(Ru(phen)_(3))〜(2+)与ss-DNA和ds-DNA的相互作用。力的测量结果表明(Ru(phen)_(3))〜(2+)与ss-DNA之间的相互作用很小,这归因于(Ru(phen)_(3))的静电结合〜(2+)到ss-DNA表面。对于ds-DNA,据认为存在强相互作用,这是由于(Ru(phen)_(3))〜(2+)与ds-DNA的缔合或插入。

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