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Nuclei Isolation from Adult Mouse Kidney for Single-Nucleus RNA-Sequencing
JoVE Journal
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Nuclei Isolation from Adult Mouse Kidney for Single-Nucleus RNA-Sequencing

Nuclei Isolation from Adult Mouse Kidney for Single-Nucleus RNA-Sequencing

DOI:
10.3791/62901-v

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06:00 min

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September 20, 2021

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, , , , , ,
1Department of Nephrology and Medical Intensive Care Medicine,Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 2Molecular and Translational Kidney Research,Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 3Berlin Institute of Medical Systems Biology (BIMSB), Systems Biology of Gene Regulatory Elements,Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 4Berlin Institute of Medical Systems Biology (BIMSB),Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC)

Bölümler

  • 00:04Introduction
  • 00:41Tissue Preparation
  • 01:41Tissue Homogenization and Cell Lysis
  • 03:11Nuclei Purification
  • 04:40Results: Single‐Nucleus RNA‐Sequencing Using Nuclei Isolated from Adult Mouse Kidney
  • 05:30Conclusion

Özet

Otomatik Çeviri

Otomatik Çeviri

Here, we present a protocol to isolate high-quality nuclei from frozen mouse kidneys that improve the representation of medullary kidney cell types and avoids the gene expression artifacts from enzymatic tissue dissociation.

Etiketler

Nuclei IsolationSingle-nucleus RNA-sequencingCortexMedullaRNA StabilizationNuclei Lysis BufferCell LysisCell Straining

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