FLUORESCENCE RESONANCE ENERGY TRANSFER DYE NUCLEOTIDE TERMINATORS: A NEW SYNTHETIC APPROACH FOR HIGH-THROUGHOUT DNA SEQUENCING

Abstract

Fluorescence resonance energy transfer (FRET) based dye-nucleotide terminators (10–13) were designed, synthesized, and formulated with Thermo Sequenase™ II DNA polymerase into a robust kit for high throughput DNA sequencing. The key energy transfer (ET) rigid and linear linker (2), required for the syntheses of energy transfer cassettes (6–9) was synthesized via Heck coupling reaction on t-Boc-L-4-iodo-phenylalanine (1) with N-TFA-propargylamine.

ACKNOWLEDGMENTS

The authors wish to thank Inna Livshin, Jim Holecek for initial HPLC experiments, Drs. Mark McDougall and C. Y. Chen for Mass and fluorescence spectral analysis.