Sensitive and powerful single-cell RNA sequencing using mcSCRB-seq

• DOI: 10.1038/s41467-018-05347-6
• Publication Date: 2018-07-20T14:52:33Z
• Authors: Johannes W. Bagnoli; Christoph Ziegenhain; Aleksandar Janjic; Lucas E. Wange; Beate Vieth; Swati Parekh; Johanna Geuder; Ines Hellmann; Wolfgang Enard
• Keywords: RNA-Seq

Abstract Single-cell RNA sequencing (scRNA-seq) has emerged as a central genome-wide method to characterize cellular identities and processes. Consequently, improving its sensitivity, flexibility, cost-efficiency can advance many research questions. Among the flexible plate-based methods, single-cell barcoding (SCRB-seq) is highly sensitive efficient. Here, we systematically evaluate experimental conditions of this protocol find that adding polyethylene glycol considerably increases sensitivity by enhancing cDNA synthesis. Furthermore, using Terra polymerase efficiency due more even amplification requires less libraries. We combined these other improvements develop scRNA-seq library call molecular crowding SCRB-seq (mcSCRB-seq), which show be one most sensitive, efficient, methods date.