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Detection of DNA hybridization by various spectroscopic methods using the copper tetraphenylporphyrin complex as a probe
Authors:Hang Gong  Changqun Cai  Ying Ma  Xiaoming Chen
Institution:1. Key Laboratory of Environmentally Friendly Chemistry and Applications of Ministry of Education, College of Chemistry, Xiangtan University, Xiangtan, Hunan, 411105, China
Abstract:We are presenting new and highly sensitive hybridization assays. They are based on various spectroscopic methods including resonance light scattering, circular dichroism, ultraviolet spectra and fluorescence spectra, as well as atomic force microscopy, and relies on the interaction of the Cu(II), Ni(II), Mg(II), Co(II), Cd(II), and Zn(II) complexes, respectively, of tetraphenylporphyrin (TPP) with double-strand DNA (dsDNA) and single strand DNA (ssDNA). The interaction results in amplified resonance light scattering (RLS) signals and enables the detection of hybridization without the need for labeling DNA. The RLS signals are strongest in case of the Cu (II)-TPP complex which therefore was selected as the probe. The technique is simple, robust, accurate, and can be completed in less than one hour.
Figure
RLS spectra of bindings of P1 and P1≈T1 with Cu (II)-TPP. The RLS intensitities of Cu (II)-TPP, P1 and P1≈T1 are weak. When P1≈T1 is mixed with Cu (II)-TPP, several enhanced RLS peaks can be observed at 362?nm, 410?nm and 471?nm(Curve 4), this enhancement is very significant, it indicate that a complex of Cu (II)-TPP-P1≈T1 is formed. With the increase of the concentration of P1≈T1, the RLS intensity increased, the enhanced RLS intensity is proportional to the concentration of P1≈T1 in an appropriate range (e.g.0.2–1.0?×?10?7?mol?L?1) ( Curve 5–7), so this method could be applied to determine oligonucleotide conveniently.
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