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 affected by anomalous migration times due to specific interactions of the DNA with the gel matrix as well as conformational differences in the DNA due to sequence heterogeneity. These anomalous migration times are strongly dependent on the choice of gel matrix. For example, the anomalous migration times that are observed in a 1 kilobase standard DNA ladder can be minimized using nongel hydroxyethylcellulose. In addition, the peak resolution can be increased and the anomalous migration can be reduced by the addition of the intercalating dye, ethidium bromide. However, in the case of the D1S80 allelic ladder, some of the DNA fragments 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.
Atha
D H
DH
Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA. donald.atha@nist.gov
D004247:DNA / Q000592:standards*; D019075:Electrophoresis, Capillary / Q000379:methods; D004996:Ethidium; D011108:Polymers; D012015:Reference Standards
D011108:Polymers; D004247:DNA; D004996:Ethidium