title: BrAD-seq: Breath Adapter Directional sequencing: a streamlined, ultra-simple and fast library preparation protocol for strand specific mRNA library construction.
abstract: Next Generation Sequencing (NGS) is driving rapid advancement in biological understanding and RNA-sequencing (RNA-seq) has become an indispensable tool for biology and medicine. There is a growing need for access to these technologies although preparation of NGS libraries remains a bottleneck to wider adoption. Here we report a novel method for the production of strand specific RNA-seq libraries utilizing the terminal breathing of double-stranded cDNA to capture and incorporate a sequencing adapter. Breath Adapter Directional sequencing (BrAD-seq) reduces sample handling and requires far fewer enzymatic steps than most available methods to produce high quality strand-specific RNA-seq libraries. The method we present is optimized for 3-prime Digital Gene Expression (DGE) libraries and can easily extend to full transcript coverage shotgun (SHO) type strand-specific libraries and is modularized to accommodate a diversity of RNA and DNA input materials. BrAD-seq offers a highly streamlined and inexpensive option for RNA-seq libraries.
authors.affiliation: Department of Plant Biology, University of California, Davis Davis, CA, USA.
authors.forename: Brad T
authors.initials: BT
authors.lastname: Townsley
authors.affiliation: Department of Plant Biology, University of California, Davis Davis, CA, USA.
authors.forename: Michael F
authors.initials: MF
authors.lastname: Covington
authors.affiliation: Department of Plant Biology, University of California, Davis Davis, CA, USA.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Ichihashi
authors.affiliation: Department of Plant Biology, University of California, Davis Davis, CA, USA.
authors.forename: Kristina
authors.initials: K
authors.lastname: Zumstein
authors.affiliation: Department of Plant Biology, University of California, Davis Davis, CA, USA.
authors.forename: Neelima R
authors.initials: NR
authors.lastname: Sinha
keywords: Bioinformatics; BrAD-seq; Illumina; NGS; RNA-seq libraries; strand-specific sequencing