NUCLEOSIDE 5'-TRIPHOSPHATE FLUORESCENT ANALOGS IN NONRADIOACTIVE METHOD FOR STUDY OF NUCLEIC-ACIDS

The 2'-deoxyuridine 5'-triphosphate analogs carrying substituents at C5 atom containing fluorescein and rhodamine type of fluorescent residues were synthesized. Avian myeloblastosis viral reverse transcriptase, rat liver DNA polymerase beta, calf thymus terminal deoxynucleotidyl transferase, and the E. coli DNA polymerase I Klenow fragment incorporated the 2'-deoxyribonucleoside 5'-triphosphate residue carrying the fluorescent label on the 3'-end of the oligonucleotide. This oligonucleotide then served as a primer for the synthesis of the (-)-strand of the M13 mp10 phage. Fluorescently labelled template-primer complexes were utilized in determinations of the primary structure of DNA.