Assaying mRNA deadenylation in vitro

Abstract

Deadenylation is the removal of poly(A) tails from mRNA. This chapter presents two methods to assay deadenylation in vitro. The fi rst is a quick and quantitative assay for the degradation of radiolabeled poly(A) that can easily be adapted to be used for many different enzymes. The second method uses an extract from Drosophila embryos to catalyze the deadenylation of an RNA dependent on a specifi c sequence that also directs deadenylation in vivo.