Sequencing of First-strand cDNA Library Reveals Full-length Transcriptomes

We developed a new method of preparing libraries for strand-specific RNA sequencing (ssRNA-Seq). It employs Direct Ligation of Adaptors to First-strand cDNA (DLAF). We compared ssRNA-Seq libraries prepared using either the DLAF and dUTP methods from mouse embryonic stem cells (mES) and libraries were sequenced from one end or both ends. We also conducted a comparison of ssRNA-Seq libraries prepared using DLAF and ScriptSeq v2 kit (Epicenter) from mouse embryonic cortex (mECx). Overall design: RNA was isolated using either Trizol or Qiagen Rneasy kit. rRNA is depleted using Eukaryote Ribominus v2 kit and libraries were prepared using one of the methods.