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DNA shape and separation efficiency in polymer media: a computerized method based on electrophoretic mobility data

D Tietz1, A Chrambach

  • 1Section on Macromolecular Analysis, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD.

Electrophoresis
|March 1, 1993
PubMed
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The ELPHOFIT program now measures polymer sieving capacity, revealing how DNA size and separation efficiency change in agarose and polyacrylamide solutions and gels. This helps understand molecular sieving in polymer media.

Area of Science:

  • Polymer Science
  • Molecular Biology
  • Biophysics

Background:

  • Ferguson plots analyze polymer solutions and molecular properties.
  • Understanding molecular sieving is crucial for separation techniques.

Purpose of the Study:

  • To extend the ELPHOFIT program for evaluating molecular sieving capacity.
  • To assess DNA shape and separation efficiency in various polymer media.

Main Methods:

  • Utilized the extended ELPHOFIT program (version 2.2).
  • Analyzed data from previous literature on DNA in agarose and polyacrylamide solutions/gels.
  • Evaluated Ferguson plots (log-mobility vs. polymer concentration).

Main Results:

  • The program quantifies the equivalent spherical size of migrating DNA.

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  • A steep, hyperbolic decline in equivalent sphere radius was observed in uncrosslinked polyacrylamide solutions.
  • Polyacrylamide gels demonstrated higher separation efficiency than solutions; agarose solutions excelled for DNA < 1 kb.

    • Conclusions:

    • ELPHOFIT 2.2 provides a measure of molecular sieving capacity.
    • DNA size and separation efficiency are significantly influenced by polymer concentration and type.
    • The program aids in characterizing DNA behavior in different polymer matrices.