Miniaturization of Smart-seq2 for Single-Cell and Single-Nucleus RNA Sequencing
This protocol presents a plate-based workflow to perform RNA sequencing analysis of single cells/nuclei using Smart-seq2. We describe (1) the dissociation procedures for cell/nucleus isolation from the mouse brain and human organoids, (2) the flow sorting of single cells/nuclei into 384-well plates,...
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Published in | STAR protocols Vol. 1; no. 2; p. 100081 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Elsevier Inc
18.09.2020
Elsevier |
Subjects | |
Online Access | Get full text |
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Summary: | This protocol presents a plate-based workflow to perform RNA sequencing analysis of single cells/nuclei using Smart-seq2. We describe (1) the dissociation procedures for cell/nucleus isolation from the mouse brain and human organoids, (2) the flow sorting of single cells/nuclei into 384-well plates, and (3) the preparation of libraries following miniaturization of the Smart-seq2 protocol using a liquid-handling robot. This pipeline allows for the reliable, high-throughput, and cost-effective preparation of mouse and human samples for full-length deep single-cell/nucleus RNA sequencing.
For complete details on the use and execution of this protocol, please refer to Bowers et al. (2020).
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•Isolation of cells or nuclei from the mouse brain and human forebrain organoids•Flow sorting of single cells or nuclei into 384-well plates•Miniaturization of the Smart-seq2 protocol using a liquid-handling robot•Protocol enables high-throughput full-length single-cell/nucleus RNA sequencing
This protocol presents a plate-based workflow to perform RNA sequencing analysis of single cells/nuclei using Smart-seq2. We describe (1) the dissociation procedures for cell/nucleus isolation from the mouse brain and human organoids, (2) the flow sorting of single cells/nuclei into 384-well plates, and (3) the preparation of libraries following miniaturization of the Smart-seq2 protocol using a liquid-handling robot. This pipeline allows for the reliable, high-throughput, and cost-effective preparation of mouse and human samples for full-length deep single-cell/nucleus RNA sequencing. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Lead Contact Technical Contact These authors contributed equally |
ISSN: | 2666-1667 2666-1667 |
DOI: | 10.1016/j.xpro.2020.100081 |