Analysis of DNA by capillary electrophoresis

Abstract

Recent advances in the separation of oligo DNA fragments using capillary electrophoresis (CE) were reviewed with special emphasis on the historical aspects and possible applications of the technology in the near future. In the section on single strand DNA separation, the high resolution capability of CE was demonstrated by referring to the size separation of oligo DNA in which a lmer difference in size of several hundreds base-mer can be recognized by using a polyacrylamide gel-filled capillary or high viscosity buffer. In addition, a statistical analysis of the migration time of heterogeneous oligo DNA was shown. By employing Gauss's least square method, the estimated migration time of any known sequences having no secondary struture can be calculated with probable error of less than 0.08 min. The effect of the secondary structure on separation was shown. The possibility of using these secondary structure effects on migration time for doing SSCP (single strand conformation polymorphism) was indicated. The section on double strand DNA separation focussed mainly on the application of CE. The possibility of detecting a specific gene or points mutation, or doing genetic diagnosis and paternal analysis was shown by referring to example of CE application to PCR (polymerase chain reaction) product analysis, ARMS (amplification refractory mutation system) analysis, RFLP (restriction fragment length polymorphism) analysis, and VNTR (variable number of tandem repeat) analysis. © 1994, The Japan Society for Analytical Chemistry. All rights reserved.