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Nuclei isolation, immunostaining, and Fluorescence-activated nuclei sorting (FANS) for Smart-Seq2

By Ester Kalef-Ezra, PhD, Dominic Horner, BSc, George Morrow, Vanda Knitlhoffer, Andy Goldson, Iain Maculay, Yanping Guo and Christos Proukakis, PhD
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Output Details

Description
This protocol describes the steps for extracting nuclei from human postmortem brain samples, immunofluorescence, and nuclei sorting (FANS) for single-nucleus RNA-seq using the Smart-Seq2 method. We have used it to isolate nuclei from human brain samples (such as the cingulate cortex), but it can be adapted for nuclei from different body areas, cell culture materials, and/or different species.
Identifier (DOI)
10.17504/protocols.io.j8nlk8wjxl5r/v1
Tags
  • Immunofluorescence
  • Nuclei isolation
  • scRNAseq (Single-cell RNA-seq)
Team
Team Voet
Program
Collaborative Research Network
Theme
PD Functional Genomics

Meet the Authors

  • Ester Kalef-Ezra, PhD

    Key Personnel: Team Voet, 

    University College London

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    Dominic Horner, BSc

    Key Personnel: Team Voet, 

    University College London

  • User avatar fallback logo

    George Morrow

    External Collaborator

  • User avatar fallback logo

    Vanda Knitlhoffer

    External Collaborator

  • User avatar fallback logo

    Andy Goldson

    External Collaborator

  • User avatar fallback logo

    Iain Maculay

    External Collaborator

  • User avatar fallback logo

    Yanping Guo

    External Collaborator

  • Christos Proukakis, PhD

    Co-PI (Core Leadership): Team Voet, 

    University College London

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Aligning Science Across Parkinson's
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