Separation of DNA by capillary electrophoresis

Methods Mol Biol. 2008:384:415-29. doi: 10.1007/978-1-59745-376-9_15.

Abstract

This chapter reports an overview of the analytical techniques used to perform genetic analysis of polymerase chain reaction products using capillary electrophoresis. Three separate but related techniques are described: the separation of native DNA with detection using fluorescent intercalating dyes, the separation of denatured DNA using fluorescently labeled primers, and the detection of single-strand conformation polymorphisms using denatured DNA separated under native conditions. The various techniques involve electrokinetic injection of the DNA onto a narrow band at the head of the column, sieving the DNA through various entangled polymer matrices, and detection via single or multichannel laser-induced fluorescence. Analytical protocols are provided, and a series of representative electropherograms are included.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA / isolation & purification*
  • DNA, Ribosomal / genetics
  • Electrophoresis, Capillary / methods*
  • Electrophoresis, Capillary / standards
  • Humans
  • Microsatellite Repeats / genetics
  • Nucleic Acid Denaturation
  • Plasmids / isolation & purification
  • Polymerase Chain Reaction
  • Polymorphism, Single-Stranded Conformational
  • Reference Standards
  • Soil Microbiology

Substances

  • DNA, Ribosomal
  • DNA