Capillary electrophoresis of DNA for molecular diagnostics

P. G. Righetti, C. Gelfi

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

A number of applications of capillary zone electrophoresis (CZE) in sieving liquid polymers (notably linear polyacrylamides and cellulose) for the analysis of polymerase chain reaction (PCR) products of clinically relevant, diagnostic DNA, are reviewed. The fields covered are: human genetics, quantitative gene dosage, microbiology and virology, forensic medicine and therapeutic DNA (notably, antisense nucleotides). Some unique, novel developments are highlighted, such as: (i) nonisocratic CZE, i.e., temperature-programmed CZE for detection of DNA point mutations; (ii) the synthesis of novel N-substituted acrylamides, offering extreme resistance to alkaline hydrolysis coupled to high hydrophilicity. In the field of denaturing gradient gel electrophoresis (DGGE), as routinely performed in gel slabs, a novel methodology is described in CZE: double-gradient DGGE. In this technique, two gradients are simultaneously applied along the migration direction: a chemical (or thermal) denaturing gradient, for partially unwinding home- and hetero-duplexes of DNA, and a porosity gradient, for recompacting diffuse bands melting over a broader range of denaturing conditions. It is thus demonstrated that chemical gradients, in addition to temperature gradients, can be easily implemented even in a capillary format.

Original languageEnglish
Pages (from-to)1709-1714
Number of pages6
JournalElectrophoresis
Volume18
Issue number10
DOIs
Publication statusPublished - 1997

Fingerprint

Capillary electrophoresis
Molecular Pathology
Capillary Electrophoresis
Electrophoresis
Denaturing Gradient Gel Electrophoresis
DNA
Acrylamides
Antisense DNA
Gels
Thermal gradients
Virology
Temperature
Forensic Medicine
Gene Dosage
Porosity
Medical Genetics
Microbiology
Hydrophobic and Hydrophilic Interactions
Point Mutation
Cellulose

Keywords

  • Capillary electrophoresis
  • Clinical chemistry
  • DNA
  • Genetic disorders

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Capillary electrophoresis of DNA for molecular diagnostics. / Righetti, P. G.; Gelfi, C.

In: Electrophoresis, Vol. 18, No. 10, 1997, p. 1709-1714.

Research output: Contribution to journalArticle

Righetti, P. G. ; Gelfi, C. / Capillary electrophoresis of DNA for molecular diagnostics. In: Electrophoresis. 1997 ; Vol. 18, No. 10. pp. 1709-1714.
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