Fluorescence detection of polyadenylation reaction through the coordination of adenosine2–coralyne–adenosine2

Abstract

A simple, sensitive, and selective-detection system for polyadenylation reaction was developed based on the fact that coralyne induces conformational change of polyadenosine [poly(A)] oligonucleotide through adenosine2–coralyne–adenosine2 coordination. Double-strand chelating dye SYBR Green I fluoresces weakly in the presence of random coil of poly(A). Coralyne enables poly(A) to change its conformation from a random coil to a folded structure, which leads to an increased fluorescence of SYBR Green I. When the 3′ end of RNA is polyadenylated with poly(A) polymerase, coralyne-induced fluorescence enhancement of the poly(A)-SYBR Green I complex is well suited for monitoring polyadenylation reaction.