Rapid hybridization of nucleic acids using isotachophoresis.

نویسندگان

  • Moran Bercovici
  • Crystal M Han
  • Joseph C Liao
  • Juan G Santiago
چکیده

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.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

On-chip Isotachophoresis and Functionalized Hydrogel Capture for Sensitive Micro-rna Detection

We integrate on-chip isotachophoresis (ITP) and photopatterned functionalized hydrogels to perform rapid high-sensitivity detection of nucleic acids. We use ITP to enhance hybridization kinetics between target microRNAs and reporter oligos and remove excess reporters with a functionalized polyacrylamide capture gel. We achieve over 4000-fold background signal reduction, ~1 pM sensitivity, 4 ord...

متن کامل

Integration of rapid DNA hybridization and capillary zone electrophoresis using bidirectional isotachophoresis.

We present a method for rapid, sequence-specific detection of multiple DNA fragments by integrating isotachophoresis (ITP) based DNA hybridization and capillary zone electrophoresis (CZE) using bidirectional ITP. Our method leverages the high preconcentration ability of ITP to accelerate slow, second-order DNA hybridization kinetics, and the high resolving power of CZE to separate and identify ...

متن کامل

Isotachophoresis with ionic spacer and two-stage separation for high sensitivity DNA hybridization assay.

We present an on-chip electrophoretic assay for rapid and high sensitivity nucleic acid (NA) detection. The assay uses isotachophoresis (ITP) to enhance NA hybridization and an ionic spacer molecule to subsequently separate reaction products. In the first stage, the probe and target focus and mix rapidly in free solution under ITP. The reaction mixture then enters a region containing a sieving ...

متن کامل

Induced nanoparticle aggregation for short nucleic acid quantification by depletion isotachophoresis.

A rapid (<20min) gel-membrane biochip platform for the detection and quantification of short nucleic acids is presented based on a sandwich assay with probe-functionalized gold nanoparticles and their separation into concentrated bands by depletion-generated gel isotachophoresis. The platform sequentially exploits the enrichment and depletion phenomena of an ion-selective cation-exchange membra...

متن کامل

Integrated printed circuit board device for cell lysis and nucleic acid extraction.

Preparation of raw, untreated biological samples remains a major challenge in microfluidics. We present a novel microfluidic device based on the integration of printed circuit boards and an isotachophoresis assay for sample preparation of nucleic acids from biological samples. The device has integrated resistive heaters and temperature sensors as well as a 70 μm × 300 μm × 3.7 cm microfluidic c...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Proceedings of the National Academy of Sciences of the United States of America

دوره 109 28  شماره 

صفحات  -

تاریخ انتشار 2012