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Resistive-pulse sensing effectively detects large DNA molecules using a nanopore. Higher voltage increases detection frequency but shortens DNA translocation events.

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Area of Science:

  • Nanotechnology
  • Biophysics
  • Molecular Biology

Background:

  • Resistive-pulse sensing is a technique for detecting molecules passing through a nanopore.
  • Large DNA molecules present unique challenges for nanopore sensing due to their size.

Purpose of the Study:

  • To investigate the resistive-pulse sensing of large single-stranded and double-stranded DNA using a conically shaped nanopore.
  • To analyze the relationship between DNA concentration, applied voltage, and translocation event characteristics.

Main Methods:

  • Utilized a track-etched polycarbonate membrane with a conically shaped nanopore (40 nm tip, 1.5 µm base).
  • Employed resistive-pulse (stochastic sensing) by applying a transmembrane potential and monitoring ion current.
  • Electrophoretically drove single-stranded phage DNA through the nanopore and analyzed translocation events.

Main Results:

  • Phage DNA translocation events appeared as transient current blocks.
  • Event frequency scaled linearly with DNA concentration and applied transmembrane potential.
  • Increased applied potential reduced the duration of current-block events.
  • Double-stranded plasmid DNA, too large for the nanopore tip, caused shorter current blocks, likely due to pore-tip collisions.

Conclusions:

  • Resistive-pulse sensing is viable for detecting large DNAs with a specifically designed nanopore.
  • Translocation event characteristics are sensitive to DNA size, concentration, and applied voltage.
  • Nanopore geometry and DNA size critically influence translocation behavior and sensing outcomes.