CAPILLARY ELECTROPHORESIS OF POLYMERASE CHAIN REACTION-AMPLIFIED DNA USING FLUORESCENCE DETECTION WITH AN INTERCALATING DYE

The application of the fluorescent intercalating dye, YO-PRO-1 to the analysis of polymerase chain reaction-generated DNA fragments from 120 to 400 base pairs in size was investigated. Analysis of samples was performed using a non-gel sieving buffer containing Tris-borate, hydroxyethyl cellulose, and the intercalating dye. Minimum detectable concentrations were less than 500 pg/ml DNA. For samples requiring resolution of four base pairs or less, it was necessary to add a second intercalating agent, ethidium bromide to the buffer. Using this procedure, a number of loci of interest in genetic typing were examined.