PEN-seq for identifying novel non-coding RNAs and their expression in cultured cells

Small RNAs between 20-500nt were least explored. Here we used RNase H-based rRNA depletion and a serial of strategies to developed PEN-seq method for identifying novel non-coding RNAs in total RNAs. Overall design: Total or subcellular RNA were ligated to small RNA-seq adapors, followed by rRNA and snRNA depletion. After reverse transcription and library amplification, inserts between 20-500bp were size selected and subjected to Illumina sequencing.