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用于单细胞和单细胞核RNA测序的Smart-seq2的小型化

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

作者信息

Jaeger Baptiste N, Yángüez Emilio, Gesuita Lorenzo, Denoth-Lippuner Annina, Kruse Merit, Karayannis Theofanis, Jessberger Sebastian

机构信息

Laboratory of Neural Plasticity, Faculties of Medicine and Science, Brain Research Institute, University of Zurich, 8057 Zurich, Switzerland.

Functional Genomic Center Zurich, ETH and University of Zurich, Zurich, Switzerland.

出版信息

STAR Protoc. 2020 Sep 18;1(2):100081. doi: 10.1016/j.xpro.2020.100081.

DOI:10.1016/j.xpro.2020.100081
PMID:33000004
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7501729/
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).

摘要

本方案介绍了一种基于平板的工作流程，用于使用Smart-seq2对单细胞/细胞核进行RNA测序分析。我们描述了(1)从小鼠脑和人类类器官中分离细胞/细胞核的解离程序，(2)将单细胞/细胞核流式分选到384孔板中，以及(3)使用液体处理机器人对Smart-seq2方案进行小型化后制备文库。该流程能够可靠、高通量且经济高效地制备用于全长深度单细胞/细胞核RNA测序的小鼠和人类样本。有关本方案使用和执行的完整详细信息，请参阅Bowers等人(2020年)的文献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/655a/7580229/8016e0417224/gr8.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/655a/7580229/1f7d8f35cf74/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/655a/7580229/24f5851ee6ac/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/655a/7580229/6d7c7b94bbcf/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/655a/7580229/2fe3d65e8b4d/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/655a/7580229/aace65796012/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/655a/7580229/064a86479cc9/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/655a/7580229/e15741f70ea1/gr6.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/655a/7580229/8016e0417224/gr8.jpg

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