high efficiency and multiplexable Strand specificity RNA sequencing by Double Adapter Directional Capture
The quantification of transcriptome through RNA sequencing (RNA-seq) has widely been used to characterize all patterns of genes in a certain period. However, although many experimental protocols have recently been developed, RNA-seq is still a challenging technology in application. It is necessary to develop an easy and versatile RNA-seq method available in a wide range from single cell RNA to bulk RNA. Here we introduce a novel RNA-seq method, Double Adapter Directional Capture sequencing (DADCSeq), by directly adding adaptor at both termini of library respectively in the steps of reverse transcription and second strand synthesis, which can not only produce high quality strand specificity full-length RNA-seq library but also reduces cockamamie steps of library constructions with less time consuming. The DADCSeq was also optimized for prokaryote such as Mycobacterium smegmatis, a model bacterium for Mycobacterium tuberculosis, and can detect more genes compared to traditional methods. Taken together the DADCSeq offers a greatly simplified and cost-effective protocol with high repeatable strand specificity RNA-seq results for a diversity of RNA input materials from prokaryote to eucaryote RNA-seq libraries. Overall design: To develop an easy and versatile RNA-seq method available in RNA library