Fluorescence resonance energy transfer terminators for DNA sequencing

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.