Sequencing of antisense DNA analogues by capillary gel electrophoresis with laser-induced fluorescence detection.
A method using capillary gel electrophoresis with laser-induced fluorescence detection is described which permits complete sequence determination of antisense DNA analogues of unknown sequence. This method, originally created as a tool to confirm the sequence of antisense oligonucleotides being developed as therapeutic drugs, utilizes data collected under a range of experimental conditions described by the Ogston model as applied to gel electrophoresis. A linear relationship independent of experimental conditions between the relative electrophoretic migration time and the oligonucleotide base number was observed and is shown to be consistent with a simplified version of this model and can be used to facilitate the sequence determination.
Belenky
A
A
Hybridon Inc., Worcester, MA 01605, USA.
Smisek
D L
DL
Cohen
A S
AS
D001483:Base Sequence; D002197:Capillary Action; D011088:DNA Ligases / Q000378:metabolism; D016373:DNA, Antisense / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005456:Fluorescent Dyes; D007834:Lasers; D008433:Mathematics; D008969:Molecular Sequence Data; D017421:Sequence Analysis / Q000379:methods*; D014466:Ultraviolet Rays
D016373:DNA, Antisense; D005456:Fluorescent Dyes; D011088:DNA Ligases