Rapid hybridization of nucleic acids using isotachophoresis

被引:79
|
作者
Bercovici, Moran [1 ]
Han, Crystal M.
Liao, Joseph C. [1 ]
Santiago, Juan G. [1 ]
机构
[1] Stanford Univ, Dept Urol, Stanford, CA 94305 USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
hybridization kinetics; DNA; RNA; molecular beacons; electrophoresis; DNA RENATURATION RATES; MOLECULAR BEACONS; PATHOGEN DETECTION; TEMPERATURE; PROBES; ACCELERATION; SPECIFICITY; KINETICS;
D O I
10.1073/pnas.1205004109
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页码:11127 / 11132
页数:6
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