Design and synthesis of fluorescence-labeled nucleotide with a cleavable aryl-triazene linker for DNA sequencing by synthesis.

Abstract

A novel acid-cleavable triazene linker was synthesized and then reacted with modified 2'-deoxyuridine triphosphate (dUTP), followed by Cy3 NHS ester, the final product serves as an excellent reversible terminator for DNA sequencing by synthesis (DNA SBS). The synthesized dye-labeled terminator incorporated into DNA strand faithfully in a DNA-elongation, and the fluorophore incorporated into DNA strands was cleaved completely under weak acidic conditions within short time. Further the first cleaved product can be incorporated with 100% efficiency for the second time. These preliminary evaluations show that the triazene reversible terminator has a great potential value in DNA sequencing.