U. Vincent et al., QUANTITATION OF POLYMERASE CHAIN REACTION-AMPLIFIED DNA FRAGMENTS BY CAPILLARY ELECTROPHORESIS AND LASER-INDUCED FLUORESCENCE DETECTION, Electrophoresis, 17(3), 1996, pp. 512-517
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.