Abstract
This chapter describes the apparatus, gel fabrication, and biochemical and data acquisition and analysis methods needed to perform fluorescence-based automated DNA sequence analysis by capillary gel electrophoresis. The procedures for the production of gel-filled capillaries are simple but can prove to be a problem if reagent quality and storage procedures are not carefully monitored. Reproducible methods and freshly prepared reagents should be used to ensure consistent results. A major problem often encountered with capillary gels is the formation of bubbles within the capillary tube that lead to a loss of resolving power or a complete failure to elute samples. The formation of bubbles in the capillary during polymerization probably originates either from the outgassing of the solvent or from a shrinkage of the polymer solution as it polymerizes.
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