5] Fluorescent nucleotides: Synthesis and characterization

Abstract

Publisher Summary The purpose of this chapter is to provide information on the fluorescent nucleotides. A proper characterization of fluorescent nucleotides requires reporting of the absorption spectrum and molar extinction coefficient. Molar extinction coefficients are useful because the exact mass of many nucleotide preparations is not known. Fluorescent nucleotides are widely used to study the structure and function of proteins and nucleic acids. With the advent of methods to study macromolecules at the single-molecule level with fluorescence imaging techniques, and the advent of two-photon excitation technologies, the interest in new nucleofide derivatives with appropriate fluorescent properties has increased. Basic information about the general approaches to synthesis such as reagent preparation, monitoring of synthetic reactions, chromatographic methods, spectral characterization, and compositional analysis are provided in the chapter to prepare new compounds. The general classes of nucleotide derivatives are also summarized and general synthetic routes are described in the chapter. Specific protocols are provided only when the synthesis demonstrates a route to a general class of compounds.