CHARACTERIZATION OF FLUORESCENT NUCLEOSIDE TRIPHOSPHATES BY CAPILLARYELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE DETECTION - ACTION OFALKALINE-PHOSPHATASE AND DNA-POLYMERASE

A method of analysis of fluor-labeled nucleoside triphosphates based o n alkaline phosphatase-catalyzed sequential cleavage of phosphate grou ps with monitoring of all fluorescent species by capillary electrophor esis with laser-induced fluorescence detection is presented. The metho d allows determination of the purity of the triphosphate samples as we ll as the relative amounts of the lower phosphate contaminants. The ab ility of one of the fluor-labeled nucleoside triphosphates to serve as polymerase substrate was verified by labeling DNA restriction fragmen ts by the method of filling recessed 3'-ends using DNA polymerase Klen ow fragment. (C) 1996 Academic Press, Inc.