REACTION OF FLUORESCAMINE WITH NITROGENOU S BASES IN NUCLEOTIDES AND SINGLE-STRANDED SYNTHETIC AND NATURAL OLIGONUCLEOTIDES

Reactions of fluorescamine with nucleoside bases have been studied. Fl uorescamine reacts with NH groups of dA, dG, dC, and the characteristi c pyrrholinone specter with maximum at 490 nm appears. Fluorescamine d oes not react with dU and dT. The reaction is pH-dependent, but not li nked to protonization of aminogroups. pK values for dA, dG, dC are res pectively 7.3, 8.4, 8.7. At pH greater than 10 or lower than 3 the flu orescence decreases clue to formation of non-fluorescent fluorescamine derivates. The average fluorescence jield of pyrrholinone (the produc t of reaction of fluorescamine with denatured DNA and single-stranded polymers containing dA) is 2.7-3.5 fold higher as compared to mixture of individual nucleotides. The studied reaction can be used sequencing of nucleic acids.