Fluorescence resonance energy transfer terminators for DNA sequencing

Abstract A four-colour set of fluorescence resonance energy transfer dideoxy nucleotide terminators ( 10 – 13 ), have been synthesised using a rigid and linear tri-functional molecule ( 2 ), synthesised via Heck coupling reaction of t -Boc- l -4-iodophenylalanine ( 1 ) with N -TFA-propargylamine. Evaluation of the terminators in DNA sequencing reactions, in combination with Thermo Sequenase™ II DNA polymerase, demonstrated them to be excellent reagents for high-throughput DNA sequencing.