DNA sequencing by capillary electrophoresis with a hydroxyethylcellulose sieving buffer.

Abstract

Capillary electrophoresis (CE) is capable of rapid, high-resolution separations of DNA. The technique has been adopted for both ssDNA and dsDNA applications. In order to make CE more convenient and cost-effective, replaceable sieving buffers have recently been developed. For DNA sequencing, the most successful of these replaceable buffers have been carefully polymerized and purified viscous linear polyacrylamide solutions. However, the hazards of acrylamide are well documented, and the care required to prepare the appropriate molecular weight polymer make this approach less than ideal. We report use of a replaceable sieving buffer suitable for DNA sequencing by CE that is easy to prepare and uses commercially available, non-toxic hydroxyethylcellulose.