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Low-input Nucleus Isolation and Multiplexing with Barcoded Antibodies of Mouse Sympathetic Ganglia for Single-nucleus RNA Sequencing
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Low-input Nucleus Isolation and Multiplexing with Barcoded Antibodies of Mouse Sympathetic Ganglia for Single-nucleus RNA Sequencing

Low-input Nucleus Isolation and Multiplexing with Barcoded Antibodies of Mouse Sympathetic Ganglia for Single-nucleus RNA Sequencing

DOI:
10.3791/63397-v

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10:44 min

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March 23, 2022

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, , , , , , ,
1Department of Anatomy & Embryology,Leiden University Medical Center, 2Department of Cardiology,Leiden University Medical Center, 3Department of Medical Statistics and Bioinformatics,Leiden University Medical Center

Capitoli

  • 00:05Introduction
  • 00:59Dissection of Adult Mouse Superior Cervical Ganglia
  • 02:12Dissection of Adult Mouse Stellate Ganglia
  • 02:57Isolation and Cryopreservation of Mouse Ganglionic Cells
  • 04:56Nucleus Isolation
  • 06:50Nucleus Barcoding with Hashtag Oligos and Multiplexing
  • 08:24Results: Quality Control Analysis of Single-Nucleus cDNA Library Preparation and snRNA-Sequencing
  • 09:55Conclusion

Summary

Traduzione automatica

Traduzione automatica

This protocol describes the detailed, low-input sample preparation for single-nucleus sequencing, including the dissection of mouse superior cervical and stellate ganglia, cell dissociation, cryopreservation, nucleus isolation, and hashtag barcoding.

Tags

Keywords: Single-nucleus RNA SequencingCardiac Sympathetic NeuronsDroplet-based SequencingHashtag OligosMultiplexingSuperior Cervical GangliaStellate GangliaTrypsin Digestion

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