QUANTITATION OF POLYMERASE CHAIN REACTION-AMPLIFIED DNA FRAGMENTS BY CAPILLARY ELECTROPHORESIS AND LASER-INDUCED FLUORESCENCE DETECTION

Quantitation of DNA fragments amplified by polymerase chain reaction ( PCR) is needed for the determination of target DNA in molecular biolog y. Capillary electrophoresis in entangled polymer solution coupled to laser-induced fluorescence detection was assessed as an alternative te chnique to conventional slab gel methods to monitor competitive PCR, w hich consists of amplifying an internal standard fragment under the sa me conditions as the target fragment. The fluorescence signal was gene rated either through end-labeling of the fragments using 5'-fluorescei n-labeled primers or through intercalation of ethidium bromide along t he double strand. It is shown that the more accurate and reliable resu lts were obtained using this latter pathway.