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Steady-state, Pre-steady-state, and Single-turnover Kinetic Measurement for DNA Glycosylase Activity
Published on: August 19, 2013
A device for performing lateral conductance measurements on individual double-stranded DNA molecules
Laurent D Menard1, Chad E Mair, Michael E Woodson
1Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
ACS Nano
|September 7, 2012
Summary
This study presents a nanofluidic device that electrically monitors DNA translocation through nanochannels. The device detects DNA by measuring changes in ionic conductance at a nanochannel intersection, enabling real-time analysis.
Area of Science:
- Nanotechnology
- Molecular Biology
- Biophysics
Background:
- Monitoring DNA translocation is crucial for genomics and diagnostics.
- Existing methods for DNA analysis can be complex and time-consuming.
- Nanofluidic devices offer potential for high-throughput, sensitive molecular detection.
Purpose of the Study:
- To develop and characterize a nanofluidic device for electrical monitoring of DNA translocation.
- To investigate the effect of DNA passage on ionic conductance in a nanochannel intersection.
- To compare different device geometries for optimal DNA detection.
Main Methods:
- Fabrication of a nanofluidic device with intersecting nanochannels.
- Electrokinetic manipulation of DNA molecules through the nanochannel.
- Electrical monitoring of ionic conductance changes in the transverse nanochannel.
- Simultaneous optical and electrical measurements for validation.
Main Results:
- A transient current response was observed when DNA translocated through the nanochannel intersection.
- The ionic current was enhanced by 5-25% in 1 M KCl solutions.
- Different device geometries exhibited distinct DNA transport dynamics.
- Optical and electrical monitoring confirmed the origin of signals from DNA transport.
Conclusions:
- The developed nanofluidic device provides an effective method for electrically monitoring DNA translocation.
- The device's sensitivity to DNA passage through the nanochannel intersection is demonstrated.
- This technology holds promise for label-free, real-time DNA analysis applications.
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