The effect of surface probe density on DNA hybridization
The effect of surface probe density on DNA hybridization
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DOI：
10.1093/nar/29.24.5163
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发表时间：
2001-12-15
影响因子：
14.9
通讯作者：
Georgiadis, RM
中科院分区：
文献类型：
--
作者：
Peterson, AW;Heaton, RJ;Georgiadis, RM
DNA互补链的杂交是所有基于微阵列的DNA变异分析技术的基本原理。在本文中，我们研究如何探针固定在表面，特别是探针密度，影响动力学的目标捕获使用表面等离子体共振（SPR）光谱，原位标记的光学方法。探针密度通过改变固定条件来控制，包括溶液离子强度、界面静电势以及是否使用双链体或单链寡核苷酸。独立的探针固定化策略是使用，我们发现，DNA膜等探针密度表现出可重复的效率和可重复的动力学探针/靶杂交。然而，杂交在双链体形成的效率和靶捕获的动力学方面强烈依赖于探针密度。我们建议，探针密度效应可以解释所观察到的目标捕获率的变化，这在以前被归因于热力学效应。
The hybridization of complementary strands of DNA is the underlying principle of all microarray-based techniques for the analysis of DNA variation. In this paper, we study how probe immobilization at surfaces, specifically probe density, influences the kinetics of target capture using surface plasmon resonance (SPR) spectroscopy, an in situ label-free optical method. Probe density is controlled by varying immobilization conditions, including solution ionic strength, interfacial electrostatic potential and whether duplex or single stranded oligonucleotides are used. Independent of which probe immobilization strategy is used, we find that DNA films of equal probe density exhibit reproducible efficiencies and reproducible kinetics for probe/target hybridization. However, hybridization depends strongly on probe density in both the efficiency of duplex formation and the kinetics of target capture. We propose that probe density effects may account for the observed variation in target-capture rates, which have previously been attributed to thermodynamic effects.