Fast and highly sensitive full-length single-cell RNA sequencing using FLASH-seq

Vincent Hahaut; Dinko Pavlinic; Walter Carbone; Sven Schuierer; Pierre Balmer; Mathieu Quinodoz; Magdalena Renner; Guglielmo Roma; Cameron S Cowan; Simone Picelli

doi:10.1038/s41587-022-01312-3

Fast and highly sensitive full-length single-cell RNA sequencing using FLASH-seq

Nat Biotechnol. 2022 Oct;40(10):1447-1451. doi: 10.1038/s41587-022-01312-3. Epub 2022 May 30.

Authors

Vincent Hahaut^{1

2}, Dinko Pavlinic^{1

2}, Walter Carbone³, Sven Schuierer³, Pierre Balmer^{1

2}, Mathieu Quinodoz^{1

2}, Magdalena Renner^{1

2}, Guglielmo Roma³, Cameron S Cowan^{1

2}, Simone Picelli^{4

5}

Affiliations

¹ Institute of Molecular and Clinical Ophthalmology Basel, Basel, Switzerland.
² Department of Ophthalmology, University of Basel, Basel, Switzerland.
³ Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Basel, Switzerland.
⁴ Institute of Molecular and Clinical Ophthalmology Basel, Basel, Switzerland. simone.picelli@iob.ch.
⁵ Department of Ophthalmology, University of Basel, Basel, Switzerland. simone.picelli@iob.ch.

Abstract

We present FLASH-seq (FS), a full-length single-cell RNA sequencing (scRNA-seq) method with increased sensitivity and reduced hands-on time compared to Smart-seq3. The entire FS protocol can be performed in ~4.5 hours, is simple to automate and can be easily miniaturized to decrease resource consumption. The FS protocol can also use unique molecular identifiers (UMIs) for molecule counting while displaying reduced strand-invasion artifacts. FS will be especially useful for characterizing gene expression at high resolution across multiple samples.

MeSH terms

Exome Sequencing
Gene Expression Profiling / methods
RNA* / genetics
Sequence Analysis, RNA / methods
Single-Cell Analysis* / methods

Substances

RNA