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Massively parallel digital transcriptional profiling of single cells.


ABSTRACT: Characterizing the transcriptome of individual cells is fundamental to understanding complex biological systems. We describe a droplet-based system that enables 3' mRNA counting of tens of thousands of single cells per sample. Cell encapsulation, of up to 8 samples at a time, takes place in ?6?min, with ?50% cell capture efficiency. To demonstrate the system's technical performance, we collected transcriptome data from ?250k single cells across 29 samples. We validated the sensitivity of the system and its ability to detect rare populations using cell lines and synthetic RNAs. We profiled 68k peripheral blood mononuclear cells to demonstrate the system's ability to characterize large immune populations. Finally, we used sequence variation in the transcriptome data to determine host and donor chimerism at single-cell resolution from bone marrow mononuclear cells isolated from transplant patients.

SUBMITTER: Zheng GX 

PROVIDER: S-EPMC5241818 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature

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Massively parallel digital transcriptional profiling of single cells.

Zheng Grace X Y GX   Terry Jessica M JM   Belgrader Phillip P   Ryvkin Paul P   Bent Zachary W ZW   Wilson Ryan R   Ziraldo Solongo B SB   Wheeler Tobias D TD   McDermott Geoff P GP   Zhu Junjie J   Gregory Mark T MT   Shuga Joe J   Montesclaros Luz L   Underwood Jason G JG   Masquelier Donald A DA   Nishimura Stefanie Y SY   Schnall-Levin Michael M   Wyatt Paul W PW   Hindson Christopher M CM   Bharadwaj Rajiv R   Wong Alexander A   Ness Kevin D KD   Beppu Lan W LW   Deeg H Joachim HJ   McFarland Christopher C   Loeb Keith R KR   Valente William J WJ   Ericson Nolan G NG   Stevens Emily A EA   Radich Jerald P JP   Mikkelsen Tarjei S TS   Hindson Benjamin J BJ   Bielas Jason H JH  

Nature communications 20170116


Characterizing the transcriptome of individual cells is fundamental to understanding complex biological systems. We describe a droplet-based system that enables 3' mRNA counting of tens of thousands of single cells per sample. Cell encapsulation, of up to 8 samples at a time, takes place in ∼6 min, with ∼50% cell capture efficiency. To demonstrate the system's technical performance, we collected transcriptome data from ∼250k single cells across 29 samples. We validated the sensitivity of the sys  ...[more]

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