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

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DOI:

10.3791/63397-v

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10:44 min

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March 23, 2022

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Yang Ge², Lieke van Roon, H. Sophia Chen², Ruben Methorst, Martin Paton, Marco C. DeRuiter, Szymon M. Kielbasa, Monique R. M. Jongbloed²

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

자동 번역

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.

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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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Chapters

Summary

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