Recent progress in DNA analysis by capillary electrophoresis

Pier Giorgio Righetti, Cecilia Gelfi, Maria Rosa D'Acunto

Research output: Contribution to journalArticle

Abstract

A number of recent developments in DNA analysis by capillary electrophoresis are here reviewed. They include capillary arrays for fast, parallel DNA sequencing as well as microfabricated capillary arrays. Microfluidic chips for DNA sizing and quantitation are also covered, as well as microdevices containing arrays of regular obstacles acting as size-separators during DNA migration. Screening of DNA point mutations by two much improved techniques is also reported: in one case, such mutations are detected (but only on relative short, ca. 60-70 base-long fragments) by free electrophoresis in rather acidic (pH ca. 3) buffers; in the case of single-strand chain polymorphism, an improved technique is described based on near-neutral pH buffers with mixtures of Tris/MES cations/zwitterions. When studying the behavior of inorganic and organic cations in the Debye-Hückel layer of DNA, it was found that the latter (especially a large number of Good's buffers and other zwitterions, such as His) would bind to the DNA filament not only via charge interaction, but also via additional bonds, notably hydrogen bonds, thus altering the electrophoretic (and possibly the biological) behavior of DNA molecules. However, whether or not borate ions would bind to DNA remains still an unsettled question. Finally, capillary electrophoresis was found to be instrumental in measuring fine physicochemical parameters pertaining to DNA polyelectrolytes, such as their free mobility and their translational diffusion coefficients.

Original languageEnglish
Pages (from-to)1361-1374
Number of pages14
JournalElectrophoresis
Volume23
Issue number10
DOIs
Publication statusPublished - 2002

Fingerprint

Capillary electrophoresis
Capillary Electrophoresis
DNA
Buffers
Cations
Borates
Microfluidics
Oligonucleotide Array Sequence Analysis
DNA Sequence Analysis
Point Mutation
Electrophoresis
Hydrogen
Polyelectrolytes
Separators
Polymorphism
Ions
Hydrogen bonds
Screening
Mutation

Keywords

  • DNA separations
  • DNA sequencing
  • Microchip devices
  • Point mutations
  • Review

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Recent progress in DNA analysis by capillary electrophoresis. / Righetti, Pier Giorgio; Gelfi, Cecilia; D'Acunto, Maria Rosa.

In: Electrophoresis, Vol. 23, No. 10, 2002, p. 1361-1374.

Research output: Contribution to journalArticle

Righetti, Pier Giorgio ; Gelfi, Cecilia ; D'Acunto, Maria Rosa. / Recent progress in DNA analysis by capillary electrophoresis. In: Electrophoresis. 2002 ; Vol. 23, No. 10. pp. 1361-1374.
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