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Bio-ingénierie

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单细胞基因组测序的超高通量微流控平台

Journal JoVE
Bio-ingénierie

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Journal JoVE Bio-ingénierie

An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing

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单细胞基因组测序的超高通量微流控平台

DOI:

10.3791/57598-v

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10:00 min

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May 23, 2018

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Benjamin Demaree², Daniel Weisgerber, Freeman Lan², Adam R. Abate^2,3

¹Department of Bioengineering and Therapeutic Sciences, California Institute for Quantitative Biosciences,University of California, San Francisco, ²UC Berkeley-UCSF Graduate Program in Bioengineering,University of California, San Francisco, ³Chan Zuckerberg Biohub

Chapitres

00:05Titre
00:48Encapsulation of Cells in Agarose Microgels
02:48Breaking the Agarose Microgels
03:54Generating Barcode Droplets by Digital PCR
04:53Tagmentation of Genomic DNA in Droplets
05:48Single-cell Barcoding by Microfluidic Double Merger
08:25Results: Ultrahigh Throughput Sequencing of Single-cell Genomes
09:24Conclusion

Summary

Traduction automatique

单细胞测序揭示了生物系统中的基因型异质性, 但目前的技术缺乏对群落组成和功能的深入分析所必需的吞吐量。在这里, 我们描述了一个微流控流程的序列 > 5万单细胞基因组从不同的细胞群。

Tags

Single-cell Genome Sequencing Microfluidics High-throughput Barcoding Cell Lysis Agarose Droplet Generation Cell Encapsulation Genomic Heterogeneity Cell Suspension PBS Surfactant Syringe Pump Temperature Control Collection Oil Removal PFO

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