Rapid hybridization of nucleic acids using isotachophoresis

Moran Bercovici, Crystal M. Han, Joseph C. Liao, Juan G. Santiago

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)11127-11132
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume109
Issue number28
DOIs
StatePublished - 10 Jul 2012

Keywords

  • DNA
  • Electrophoresis
  • Hybridization kinetics
  • Molecular beacons
  • RNA

All Science Journal Classification (ASJC) codes

  • General

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