Synthesis of a fluorescent derivative of 6-N-[N-(6-aminohexyl)carbamoyl)-2′,3′-dideoxyadenosine 5′-triphosphate for detection of nucleic acids

Abstract

Treatment of 5′-O-TBDMS-2′,3′-dideoxyadenosine 2 with phenoxycarbonyltetrazole followed by N-trifluoroacetyl-1,6-diaminohexane yielded 6-N-carbamoyl derivative 4, which was further converted into the deprotected nucleoside 5. The latter compound was transformed into its triphosphate 6, which has been dye-labelled at the aliphatic amino group with the fluorescent dye JA242. The conjugate was shown to terminate DNA synthesis catalysed by terminal transferase and different DNA polymerases, allowing DNA detection by time-resolved fluorescence.