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低输入细胞核分离和多路复用小鼠交感神经节条形码抗体进行单核RNA测序
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JoVE Journal Medicine
Low-input Nucleus Isolation and Multiplexing with Barcoded Antibodies of Mouse Sympathetic Ganglia for Single-nucleus RNA Sequencing
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低输入细胞核分离和多路复用小鼠交感神经节条形码抗体进行单核RNA测序

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

Chapters

  • 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

Automatic Translation

Automatic Translation

该协议描述了用于单核测序的详细，低输入样品制备，包括小鼠上颈神经节和星状神经节的解剖，细胞解离，冷冻保存，细胞核分离和标签条形码。

Tags

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

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