Polyadenylated RNA Sampling

Abstract

Polyadenylation is a post-transcriptional modification of RNA occurring in prokaryotes, eukaryotes and organelles. However, the function and extent of bacterial polyadenylation are in marked contrast to those of eukaryotic poly(A) tails. In fact, the long poly(A) tails of eukaryotic mRNAs play an important role in their exportation to the cytoplasm and promote mRNA stability and translation, whereas the short bacterial tails facilitate RNA decay. One of the obstacles encountered by investigators studying bacterial polyadenylation is the scarcity of polyadenylated RNAs. The method described here allows reverse transcription and PCR amplification of the whole population of polyadenylated RNAs provided that the poly(A) tails are long enough to hybridize to oligo dT30 sequence. To this end utilization of exoribonucleases deficient strains may be useful.