Rapid hybridization of nucleic acids using isotachophoresis.
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.
Mechanical Engineering, Stanford University, 440 Escondido Mall, Stanford, CA 94305, USA.
Moran
M
Bercovici
Crystal M
CM
Han
Joseph C
JC
Liao
Juan G
JG
Santiago
D002021:Buffers; D004247:DNA / Q000737:chemistry; D004586:Electrophoresis / Q000379:methods; D006801:Humans; D057625:Isotachophoresis / Q000295:instrumentation / Q000379:methods*; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008956:Models, Chemical; D015233:Models, Statistical; D016011:Normal Distribution; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009696:Nucleic Acids; D012313:RNA / Q000737:chemistry; D012996:Solutions
D002021:Buffers; D009696:Nucleic Acids; D012996:Solutions; D012313:RNA; D004247:DNA