Rapid hybridization of nucleic acids using isotachophoresis
Rapid hybridization of nucleic acids using isotachophoresis
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DOI：
10.1073/pnas.1205004109
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发表时间：
2012-07-10
影响因子：
11.1
通讯作者：
Santiago, Juan G.
中科院分区：
文献类型：
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作者：
Bercovici, Moran;Han, Crystal M.;Santiago, Juan G.
我们使用等速电泳法(ITP)来控制和提高自由溶液中的核酸杂交反应的速度。我们提出了一种新的物理模型、验证实验和该分析的演示。研究了ITP下预浓缩、混合和化学反应动力学的耦合物理化学过程。我们的实验验证模型实现了ITP辅助反应动力学的封闭形式解，并揭示了一个新的特征时间尺度，它正确地预测了100 PM量级反应物的化学反应速度加快了10,000倍，并且在较低浓度下更大地提高了速度。在500 PM浓度下，我们测得的反应时间比标准二级杂交预测的反应时间要短14,000倍。该模型和方法普遍适用于涉及核酸的反应的加速，并可能适用于涉及离子反应物的广泛的反应。
We use isotachophoresis (ITP) to control and increase the rate of nucleic acid hybridization reactions in free solution. We present a new physical model, validation experiments, and demonstrations of this assay. We studied the coupled physicochemical processes of preconcentration, mixing, and chemical reaction kinetics under ITP. Our experimentally validated model enables a closed form solution for ITP-aided reaction kinetics, and reveals a new characteristic time scale which correctly predicts order 10,000-fold speed-up of chemical reaction rate for order 100 pM reactants, and greater enhancement at lower concentrations. At 500 pM concentration, we measured a reaction time which is 14,000-fold lower than that predicted for standard second-order hybridization. The model and method are generally applicable to acceleration of reactions involving nucleic acids, and may be applicable to a wide range of reactions involving ionic reactants.