PAS-seq 2: A fast and sensitive method for global profiling of polyadenylated RNAs.

Alternative polyadenylation (APA) is a widespread phenomenon in eukaryotes that contributes to regulating gene expression and generating proteomic diversity. APA plays critical roles in development and its mis-regulation has been implicated in a wide variety of human diseases, including cancer. To study APA on the transcriptome-wide level, numerous deep sequencing methods that capture 3' end of mRNAs have been developed in the past decade, but they generally require a large amount of hands-on time and/or high RNA input. Here, we introduce PAS-seq 2, a fast and sensitive method for global and quantitative profiling of polyadenylated RNAs. Compared to our original PAS-seq, this method takes less time and requires much lower total RNA input due to improvement in the reverse transcription process. PAS-seq 2 can be applied to both APA and differential gene expression analyses.