4.1 Article

Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics

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STAR PROTOCOLS
Volume 2, Issue 3, Pages -

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ELSEVIER
DOI: 10.1016/j.xpro.2021.100612

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Funding

  1. Danish National Research Foundation [141]
  2. Novo Nordisk Foundation

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Traditional single-cell analysis methods are limited in studying adipose tissues, but single-nucleus sequencing technology can overcome this issue, enabling transcriptomic profiling of the entire adipose tissue.
Lipid-filled adipocytes are incompatible with droplet-based single-cell methods, such as 10x Genomics-based technology, thus restricting droplet-based single cell analyses of adipose tissues to the stromal vascular fraction. To overcome this limitation and obtain cellular and molecular insight into adipose tissue composition and plasticity, single-nucleus sequencing-based technologies can be applied. Here, we provide an optimized protocol for nuclei isolation from mouse adipose tissues suitable for single-nucleus RNA sequencing. This allows for transcriptomic profiling of the entire adipose tissue at single-cell resolution. For complete details on the use of this protocol, please refer to Sarvari et al., 2021.

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