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

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

  • Jaeger BN Laboratory of Neural Plasticity, Faculties of Medicine and Science, Brain Research Institute, University of Zurich, 8057 Zurich, Switzerland.
  • Yángüez E Functional Genomic Center Zurich, ETH and University of Zurich, Zurich, Switzerland.
  • Gesuita L Laboratory of Neural Circuit Assembly, Faculties of Medicine and Science, Brain Research Institute, University of Zurich, 8057 Zurich, Switzerland.
  • Denoth-Lippuner A Laboratory of Neural Plasticity, Faculties of Medicine and Science, Brain Research Institute, University of Zurich, 8057 Zurich, Switzerland.
  • Kruse M Laboratory of Neural Plasticity, Faculties of Medicine and Science, Brain Research Institute, University of Zurich, 8057 Zurich, Switzerland.
  • Karayannis T Laboratory of Neural Circuit Assembly, Faculties of Medicine and Science, Brain Research Institute, University of Zurich, 8057 Zurich, Switzerland.
  • Jessberger S Laboratory of Neural Plasticity, Faculties of Medicine and Science, Brain Research Institute, University of Zurich, 8057 Zurich, Switzerland.
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  • 2020-10-01
Published in:
  • STAR protocols. - 2020
English 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).
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/79457
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