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Tissue Handling and Dissociation for Single-Cell RNA-Seq

  • Felipe A. Vieira BragaEmail author
  • Ricardo J. Miragaia
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1979)

Abstract

The starting material for all single-cell protocols is a cell suspension. The particular functions and spatial distribution of immune cells generally make them easy to isolate them from the tissues where they dwell. Here we describe tissue dissociation protocols that have been used to obtain human immune cells from lymphoid and nonlymphoid tissues to be then used as input to single-cell methods. We highlight the main factors that can influence the final quality of single-cell data, namely the stress signatures that can bias its interpretation.

Key words

Single-cell RNA sequencing Tissue processing Digestion Single-cell suspension Immune cells 

References

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Wellcome Sanger Institute, Wellcome Genome CampusHinxton, CambridgeUK

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