Related Experiment Video
Updated: Jul 8, 2026
09:43
Fine-tuning the Size and Minimizing the Noise of Solid-state Nanopores
Published on: October 31, 2013
A single-molecule nanopore device detects DNA polymerase activity with single-nucleotide resolution
Scott L Cockroft1, John Chu, Manuel Amorin
1Department of Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|January 2, 2008
Summary
Researchers observed DNA polymerase activity base-by-base at the single-molecule level. This breakthrough uses a nanopore device to monitor DNA synthesis, advancing single-molecule DNA sequencing.
Area of Science:
- Biophysics
- Molecular Biology
- Nanotechnology
Background:
- Single-molecule DNA sequencing requires monitoring DNA polymerase activity at single-nucleotide resolution.
- Current methods face challenges in achieving precise, real-time monitoring of polymerase activity.
Purpose of the Study:
- To report the first observation of base-by-base DNA polymerase activity with single-base resolution at the single-molecule level.
- To develop and characterize a novel nanopore device for this purpose.
Main Methods:
- Assembled a supramolecular nanopore device using an alpha-hemolysin protein pore in a lipid membrane.
- Threaded a DNA-PEG copolymer strand within the pore.
- Monitored ion current changes caused by single-nucleotide primer extensions and template DNA displacement.
Main Results:
- Achieved the first single-molecule, base-by-base observation of DNA polymerase activity.
- Detected up to nine consecutive single-nucleotide primer extensions with high sensitivity (
- Observed stepped changes in ion current corresponding to each nucleotide addition.
Conclusions:
- The developed nanopore device enables monitoring of DNA polymerase activity at single-nucleotide resolution.
- This represents a significant advancement for nanopore-mediated single-molecule DNA sequencing.
- The system may also be applicable to studying other biopolymer-protein interactions.
