1 April 1996 Fluorescent DNA base analogs: preparation, incorporation into oligonucleotides, and time-resolved fluorescence spectroscopy
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Abstract
The synthesis of fluorescent DNA base analogs that can replace the natural bases adenine, thymine and cytosine and their incorporation into synthetic oligodeoxynucleotides is described. The effect on the stability of such modified nucleotides like 2-aminopurine and some pteridine derivatives, is studied by thermal melting studies and comparison with the corresponding unaltered oligonucleotides. The fluorescence spectroscopic properties and several applications of these new fluorescent DNA probes are described in greater detail. Structural information on the conformation of special oligonucleotides like hairpins, junctions and bulged duplexes can be obtained from fluorescence lifetime and fluorescence depolarization data. For example the fluorescence lifetime pattern of 2-aminopurine is a sensitive indicator of DNA base pairing. As examples the structure of the oligonucleotide-linker junction in a synthetically linked oligonucleotide hairpin and the base-pairing of the first 'deoxyribozyme' are discussed. A third application uses doubly, i.e., donor and acceptor, labeled oligonucleotides to measure distances by fluorescence resonance energy transfer.
© (1996) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Remo A. Hochstrasser, "Fluorescent DNA base analogs: preparation, incorporation into oligonucleotides, and time-resolved fluorescence spectroscopy", Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); doi: 10.1117/12.237614; https://doi.org/10.1117/12.237614
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