Deadenylation is the removal of poly(A) tails from mRNA. Here, we present two methods for assaying deadenylation in vivo. The first is a method for measuring bulk poly(A) tail lengths. When combined with a block in transcription, the method can be used for measuring the rate of bulk poly(A) tail shortening. The second is an RT-PCR method to determine the poly(A) tail lengths of individual RNAs. Again in combination with a block of transcription, the method permits the rate of deadenylation of an individual RNA to be measured.