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The efficiency of fluorescence DNA labeling was estimated for four fluorescent 2′-deoxyuridine 5′-triphosphate derivatives differing in the orientation of the main dye axis, which passes through the polymethine chain, relative to the linker connecting the dye to the nucleotide. To estimate the polymerase chain reaction (PCR) rate, real-time PCR was run with two commercial hot-start DNA polymerases...
The effect of the chromophore charge on the efficiency of incorporation of fluorescent-labeled nucleotides into DNA during PCR was studied using three dUTP derivatives that contain different fluorescent labels, i.e., electroneutral, positively charged, and negatively charged Cy5 analogs. dUTP labeled with an electroneutral Cy5 analog was shown to be most efficiently incorporated into DNA when Tag...
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