Design and synthesis of fluorescence-labeled nucleotide with a cleavable azo linker for DNA sequencing

Abstract

A cleavable azo linker was synthesized and reacted with 5-(6)-carboxytetramethyl rhodamine succinimidyl ester, followed by further reactions with di(N-succinimidyl) carbonate and 5-(3-amino-1-propynyl)-2′-deoxyuridine 5′-triphosphate [dUTP(AP₃)] to obtain the terminal product dUTP-azo linker-TAMRA as a potential reversible terminator for DNA sequencing by synthesis with no need for 3′-OH blocking.