CHARACTERIZATION OF FLUORESCENT NUCLEOSIDE TRIPHOSPHATES BY CAPILLARYELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE DETECTION - ACTION OFALKALINE-PHOSPHATASE AND DNA-POLYMERASE
Ra. Evangelista et al., CHARACTERIZATION OF FLUORESCENT NUCLEOSIDE TRIPHOSPHATES BY CAPILLARYELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE DETECTION - ACTION OFALKALINE-PHOSPHATASE AND DNA-POLYMERASE, Analytical biochemistry, 235(1), 1996, pp. 89-97
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.