@article{Sallés01061995, author = {Sallés, F J and Strickland, S}, title = {Rapid and sensitive analysis of mRNA polyadenylation states by PCR.}, volume = {4}, number = {6}, pages = {317-321}, year = {1995}, abstract ={A rapid and sensitive technique is described that measures the length of the poly(A) tail on a specific mRNA within subnanogram quantities of total cellular RNA [the Poly(A) test (PAT)]. In a single-tube reaction, a poly(dT) primer is synthesized in situ on the poly(A) tail of mRNAs using oligo(dT) and DNA ligase. By modulating the annealing temperature and primer concentrations, a GC-rich adapter sequence is targeted to the 5' end of the poly(dT) primer. This ligated poly(dT)-anchor is then used to prime reverse transcription of the mRNA, yielding a library of PAT cDNAs. The length of a poly(A) tail is determined by PCR amplification using the oligo(dT)-anchor primer and a message-specific primer. Comparison of PCR products from different samples allows quantitative determination of changes in polyadenylation of a given mRNA. This technique overcomes many of the pitfalls associated with conventional poly(A) tail length assessments and should prove useful in studying a variety of processes relating to polyadenylation.}, URL = {http://genome.cshlp.org/content/4/6/317.abstract}, eprint = {http://genome.cshlp.org/content/4/6/317.full.pdf+html}, journal = {Genome Research} }