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 chemic...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2012-07, Vol.109 (28), p.11127-11132
Main Authors: Bercovici, Moran, Han, Crystal M, Liao, Joseph C, Santiago, Juan G
Format: Article
Language:English
Subjects:
Analytical models
Beacons
Biological Sciences
Buffers
Chemical reactions
DNA
DNA - chemistry
Electrophoresis - methods
Genetic hybridization
Humans
Hybridization
Isotachophoresis - instrumentation
Isotachophoresis - methods
Kinetics
Liver - metabolism
mixing
Modeling
Models, Chemical
Models, Statistical
Normal Distribution
Nucleic acid hybridization
Nucleic Acid Hybridization - methods
Nucleic Acids
Reactants
Reaction kinetics
RNA - chemistry
Solutions
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Summary: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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1205004109