Sizing DNA Using a Nanometer-Diameter Pore

Each species from bacteria to human has a distinct genetic fingerprint. Therefore, a mechanism that detects a single molecule of DNA represents the ultimate analytical tool. As a first step in the development of such a tool, we have explored using a nanometer-diameter pore, sputtered in a nanometer-...

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Bibliographic Details
Published in:Biophysical journal 2004-10, Vol.87 (4), p.2905-2911
Main Authors: Heng, Jiunn B., Ho, Chuen, Kim, Taekyung, Timp, Rolf, Aksimentiev, Aleksij, Grinkova, Yelena V., Sligar, Stephen, Schulten, Klaus, Timp, Gregory
Format: Article
Language:English
Subjects:
Biophysics
Deoxyribonucleic acid
DNA
DNA - analysis
DNA - chemistry
DNA - ultrastructure
Electrochemistry - instrumentation
Electrochemistry - methods
Electrolytes
Membranes
Molecules
Nanotechnology - instrumentation
Nanotechnology - methods
Other
Particle Size
Polymers
Porosity
Reproducibility of Results
Sensitivity and Specificity
Ultrafiltration - instrumentation
Ultrafiltration - methods
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Summary:Each species from bacteria to human has a distinct genetic fingerprint. Therefore, a mechanism that detects a single molecule of DNA represents the ultimate analytical tool. As a first step in the development of such a tool, we have explored using a nanometer-diameter pore, sputtered in a nanometer-thick inorganic membrane with a tightly focused electron beam, as a transducer that detects single molecules of DNA and produces an electrical signature of the structure. When an electric field is applied across the membrane, a DNA molecule immersed in electrolyte is attracted to the pore, blocks the current through it, and eventually translocates across the membrane as verified unequivocally by gel electrophoresis. The relationship between DNA translocation and blocking current has been established through molecular dynamics simulations. By measuring the duration and magnitude of the blocking current transient, we can discriminate single-stranded from double-stranded DNA and resolve the length of the polymer.
ISSN:0006-3495
1542-0086
DOI:10.1529/biophysj.104.041814