On-line melting of double-stranded DNA for analysis of single-stranded DNA using capillary electrophoresis

J Chromatogr B Biomed Appl. 1996 Jan 26;675(2):205-11. doi: 10.1016/0378-4347(95)00370-3.

Abstract

Capillary electrophoresis (CE) is a convenient, fast and non-radioactive method with possibilities for automatization. To analyse single-stranded DNA molecules in a more automated way, we developed a heating device to melt double-stranded DNA fragments in the capillary during electrophoresis. In this study we used this device to obtain single-stranded DNA, necessary for the detection of point mutations in DNA using the single-strand conformation polymorphism technique. Results show that double-stranded DNA molecules can be melted on-line into single-stranded DNA molecules, although not for 100%. In an attempt to find universal electrophoretic conditions for the analysis of single-stranded DNA, we investigated the influence of several parameters on the yield of single-stranded DNA molecules and on the resolution of the single-stranded DNA peaks. We demonstrate that this heating device is a technical adjustment of CE which contributes to more automated analyses of DNA fragments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry*
  • DNA / genetics
  • DNA, Single-Stranded / analysis*
  • DNA, Single-Stranded / genetics
  • Electrophoresis, Capillary / methods*
  • Genes, p53
  • Genes, ras
  • Point Mutation
  • Polymerase Chain Reaction

Substances

  • DNA, Single-Stranded
  • DNA