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DNA analysis using an electrospray scanning mobility particle sizer

S Mouradian1, J W Skogen, F D Dorman

  • 1Department of Chemistry, University of Wisconsin, Madison 53706-1396, USA.

Analytical Chemistry
|March 1, 1997
PubMed
Summary
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Electrospray-Scanning Mobility Particle Sizer (ES-SMPS) successfully analyzes DNA. This technique separates and quantifies single- and double-stranded DNA from 6.1 kDa to 300 kDa with high sensitivity.

Area of Science:

  • Analytical Chemistry
  • Biophysics
  • Molecular Biology

Background:

  • Scanning Mobility Particle Sizer (SMPS) traditionally sizes gas-phase particles by electrophoretic mobility.
  • Electrospray (ES) source addition extends SMPS analysis to the macromolecular range, enabling new applications.

Purpose of the Study:

  • To demonstrate the application of ES-SMPS for the analysis of nucleic acids, specifically DNA.
  • To evaluate the technique's ability to separate, detect, and quantify DNA molecules.

Main Methods:

  • Utilized an electrospray source coupled with a scanning mobility particle sizer (ES-SMPS).
  • Analyzed single- and double-stranded DNA molecules ranging from 6.1 kDa to 300 kDa.
  • Measured electrophoretic mobility diameters and peak areas for quantitative analysis.

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Main Results:

  • ES-SMPS successfully separated and detected DNA molecules at picomole to femtomole levels.
  • Electrophoretic mobility diameters correlated with molecular weights, and peak areas provided quantitative data.
  • No DNA fragmentation was observed; distinct apparent densities were noted for single- and double-stranded DNA.

Conclusions:

  • ES-SMPS is a viable and sensitive technique for characterizing nucleic acids.
  • The method offers fast analysis (approx. 3 min/spectrum) and distinguishes between DNA types.
  • Further optimization promises ES-SMPS as a key tool for biopolymer characterization.