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

Johannes W. Bagnoli, Christoph Ziegenhain, Aleksandar Janjic, Lucas E. Wange, Beate Vieth, Swati Parekh, Johanna Geuder, Ines Hellmann and Wolfgang Enard ()
Additional contact information
Johannes W. Bagnoli: Ludwig-Maximilians-University
Christoph Ziegenhain: Ludwig-Maximilians-University
Aleksandar Janjic: Ludwig-Maximilians-University
Lucas E. Wange: Ludwig-Maximilians-University
Beate Vieth: Ludwig-Maximilians-University
Swati Parekh: Ludwig-Maximilians-University
Johanna Geuder: Ludwig-Maximilians-University
Ines Hellmann: Ludwig-Maximilians-University
Wolfgang Enard: Ludwig-Maximilians-University

Nature Communications, 2018, vol. 9, issue 1, 1-8

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

Date: 2018
References: Add references at CitEc
Citations: View citations in EconPapers (4)

Downloads: (external link)
https://www.nature.com/articles/s41467-018-05347-6 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-05347-6

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-018-05347-6

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().