Correlation of nucleotide incorporation rate and HPLC retention parameters of substituted nucleosides

A series of 5-alkyl, alkenyl, and alkynyl substituted deoxyuridines and their triphosphate derivatives have been synthesized and studied in DNA polymerase reactions. Initial rate of incorporation of the derivatives catalyzed by Klenow fragment DNA polymerase enzyme (Escherichia coli ) was measured. The rate values expressed in relative percentage to the incorporation rate of the natural substrate dTTP can be considered as a measure of substrate specificity of the DNA polymerase enzyme.