CHARACTERIZATION OF DNA STANDARDS BY CAPILLARY-ELECTROPHORESIS

We have used capillary electrophoresis to evaluate commercial DNA size standards and have found that it can provide an efficient assessment of size. However, the accuracy of the determination is adversely affec ted by anomalous migration times due to specific interactions of the D NA with the gel matrix as well as conformational differences in the DN A due to sequence heterogeneity. These anomalous migration times are s trongly dependent on the choice of gel matrix. For example, the anomal ous migration times that are observed in a 1 kilobase standard DNA lad der can be minimized using nongel hydroxyethylcellulose. In addition, the peak resolution can be increased and the anomalous migration can b e reduced by the addition of the intercalating dye, ethidium bromide. However, in the case of the D1S80 allelic ladder, some of the DNA frag ments possess nucleotide sequences which do not interact equivalently with the dye and produce irregular migration times. These measurements yield preliminary information useful in evaluating DNA size standards which may be used for a wide range of DNA diagnostic applications.