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利用表面等离子体共振增强荧光光谱研究DNA-DNA和PNA-DNA相互作用的动力学。

Investigating the kinetics of DNA-DNA and PNA-DNA interactions using surface plasmon resonance-enhanced fluorescence spectroscopy.

作者信息

Kambhampati D, Nielsen P E, Knoll W

机构信息

Max-Planck-Institut für Polymerforschung, Postfach 3148, Ackermannweg 10, 55128 Mainz, Germany.

出版信息

Biosens Bioelectron. 2001 Dec;16(9-12):1109-18. doi: 10.1016/s0956-5663(01)00239-1.

Abstract

Plasmon surface polaritons, resonantly excited in the Kretschmann format, are used to enhance the fluorescence emission of chromophore-labeled oligonucleotides (15mers) binding to surface-attached (via biotin-streptavidin linkages) complement catcher probes. A detailed analysis of the association and dissociation kinetics as well as the affinity constants is given for a mismatch 1 hybrid, emphasizing, in particular, the experimental conditions that are required to allow for an artifact-free determination of rate constants. A first comparison between DNA- and peptide nucleic acid (PNA-) probes shows similar affinities, however, significant deviations from single-exponential kinetics predicted by a simple Langmuir model for the PNA case are found.

摘要

以Kretschmann模式共振激发的表面等离激元极化子,用于增强与表面附着(通过生物素-链霉亲和素连接)的互补捕获探针结合的发色团标记寡核苷酸(15聚体)的荧光发射。对错配1杂交体的缔合和解离动力学以及亲和常数进行了详细分析,特别强调了为无假象地测定速率常数所需的实验条件。DNA探针和肽核酸(PNA)探针的首次比较显示出相似的亲和力,然而,发现PNA的情况与简单朗缪尔模型预测的单指数动力学存在显著偏差。

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