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Miniaturization of Smart-seq2 for Single-Cell and Single-Nucleus RNA Sequencing


Jaeger, Baptiste N; Yángüez, Emilio; Gesuita, Lorenzo; Denoth-Lippuner, Annina; Kruse, Merit; Karayannis, Theofanis; Jessberger, Sebastian (2020). Miniaturization of Smart-seq2 for Single-Cell and Single-Nucleus RNA Sequencing. STAR Protocols, 1(2):100081.

Abstract

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).

Abstract

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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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Brain Research Institute
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:German
Date:18 September 2020
Deposited On:26 Jan 2021 05:37
Last Modified:01 Feb 2021 16:28
Publisher:Cell Press (Elsevier)
ISSN:2666-1667
OA Status:Gold
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.1016/j.xpro.2020.100081
PubMed ID:33000004

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