Application of capillary electrophoresis to the measurement of oligonucleotide concentration and purity over a wide dynamic range.
Synthetic oligonucleotides rarely contain 100% of the full-length sequence due, in part, to the failure sequences produced during synthesis. In this paper, a method is described for the determination of both the concentration and the purity of oligonucleotides, utilizing capillary electrophoresis with a deoxyribo-nucleoside triphosphate as an internal standard. This method is advantageous for several reasons: (a) the wide dynamic range allows for the analysis of samples without the need for dilutions; (b) a small sample size is used for analysis; (c) capillary electrophoresis is automatable which allows for high throughput; and (d) all of the samples are analyzed at the same run temperature which aids in reproducibility and consistency between runs performed at different times.
Lowery
J D
JD
DNA Diagnostics Business Unit, Bio-Rad Laboratories, Hercules, California 94547, USA.
Ugozzoli
L
L
Wallace
R B
RB
D003838:Deoxyadenine Nucleotides / Q000302:isolation & purification / Q000592:standards; D019075:Electrophoresis, Capillary / Q000379:methods*; D009841:Oligonucleotides / Q000302:isolation & purification*; D013053:Spectrophotometry
D003838:Deoxyadenine Nucleotides; D009841:Oligonucleotides; C026600:2'-deoxyadenosine triphosphate