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Lightning Fast and Highly Sensitive Full-Length Single-cell sequencing using FLASH-Seq

Hahaut, Vincent, Pavlinic, Dinko, Cowan, Cameron, Schuierer, Sven, Carbone, Walter, Roma, Guglielmo and Picelli, Simone (2022) Lightning Fast and Highly Sensitive Full-Length Single-cell sequencing using FLASH-Seq. Nature biotechnology, 40. ISSN 1087-0156

Abstract

In the last 10 years, single-cell RNA-sequencing (scRNA-seq) has undergone
exponential growth. Emulsion droplets methods1–3, such as those commercialized by 10x
Genomics, have allowed researchers to analyze tens of thousands of cells in parallel in a
robust and reproducible way. However, in contrast to SMART-based full-length
sequencing protocols4,5, these methods interrogate only the outer portion of the
transcripts and still lack the required sensitivity for analyzing comprehensively the
transcriptome of individual cells. Building upon the existing SMART-seq forerunners
protocols4,5, we developed FLASH-Seq (FS), a new scRNA-seq method which displays
greater sensitivity while decreasing incubation times and reducing the number of
processing steps compared to its predecessors. The entire FS protocol - from lysed cells
to pooled cDNA libraries - can be performed in ~4.5 hours, is automation-friendly and can
be easily miniaturized to decrease costs.

Item Type: Article
Keywords: scRNA-seq; full length transcript sequencing;
Date Deposited: 14 Jun 2022 00:45
Last Modified: 14 Jun 2022 00:45
URI: https://oak.novartis.com/id/eprint/45354

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