Comparative study of capillary zone electrophoresis and high-performance liquid chromatography in the analysis of oligonucleotides and DNA.
Capillary zone electrophoresis and high-performance liquid chromatography were compared with regard to the separation of oligonucleotides and double-stranded DNA. Both anion-exchange and reversed-phase high-performance liquid chromatography on non-porous particles are considered to be superior to capillary electrophoresis in terms of speed and selectivity in the analysis of oligonucleotides up to 30 bases in length. Moreover, reversed-phase chromatography allows the simultaneous purification of detritylated oligonucleotides with recoveries > 90%. Compared with anion-exchange chromatography, there is no need for a subsequent desaltation step because the volatile buffer system can be readily evaporated. With regard to dsDNA, however, the resolving power of capillary electrophoresis cannot be matched by anion-exchange chromatography at present. Moreover, the combined use of hydroxyethylcellulose and ethidium bromide not only yielded a separation efficiency equal to that achieved by means of gel-filled capillaries but also avoids some of their limitations such as the destruction of the gel matrix at high current densities and the bias involved in electrokinetic injection.
Oefner
P J
PJ
Department of Analytical Chemistry, Johannes-Kepler-University, Linz, Austria.
Bonn
G K
GK
Huber
C G
CG
Nathakarnkitkool
S
S
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange; D004247:DNA / Q000032:analysis*; D004586:Electrophoresis / Q000379:methods*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000032:analysis*; D016133:Polymerase Chain Reaction
D009838:Oligodeoxyribonucleotides; D004247:DNA