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

Sequencing technologies have undergone a paradigm shift from bulk to single-cell resolution in response to an evolving understanding of the role of cellular heterogeneity in biological systems. However, single-cell sequencing of large populations has been hampered by limitations in processing genome...

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Detalles Bibliográficos
Autores principales: Demaree, Benjamin, Weisgerber, Daniel, Lan, Freeman, Abate, Adam R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6101372/
https://www.ncbi.nlm.nih.gov/pubmed/29889211
http://dx.doi.org/10.3791/57598
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author Demaree, Benjamin
Weisgerber, Daniel
Lan, Freeman
Abate, Adam R.
author_facet Demaree, Benjamin
Weisgerber, Daniel
Lan, Freeman
Abate, Adam R.
author_sort Demaree, Benjamin
collection PubMed
description Sequencing technologies have undergone a paradigm shift from bulk to single-cell resolution in response to an evolving understanding of the role of cellular heterogeneity in biological systems. However, single-cell sequencing of large populations has been hampered by limitations in processing genomes for sequencing. In this paper, we describe a method for single-cell genome sequencing (SiC-seq) which uses droplet microfluidics to isolate, amplify, and barcode the genomes of single cells. Cell encapsulation in microgels allows the compartmentalized purification and tagmentation of DNA, while a microfluidic merger efficiently pairs each genome with a unique single-cell oligonucleotide barcode, allowing >50,000 single cells to be sequenced per run. The sequencing data is demultiplexed by barcode, generating groups of reads originating from single cells. As a high-throughput and low-bias method of single-cell sequencing, SiC-seq will enable a broader range of genomic studies targeted at diverse cell populations.
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spelling pubmed-61013722018-09-11 An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing Demaree, Benjamin Weisgerber, Daniel Lan, Freeman Abate, Adam R. J Vis Exp Bioengineering Sequencing technologies have undergone a paradigm shift from bulk to single-cell resolution in response to an evolving understanding of the role of cellular heterogeneity in biological systems. However, single-cell sequencing of large populations has been hampered by limitations in processing genomes for sequencing. In this paper, we describe a method for single-cell genome sequencing (SiC-seq) which uses droplet microfluidics to isolate, amplify, and barcode the genomes of single cells. Cell encapsulation in microgels allows the compartmentalized purification and tagmentation of DNA, while a microfluidic merger efficiently pairs each genome with a unique single-cell oligonucleotide barcode, allowing >50,000 single cells to be sequenced per run. The sequencing data is demultiplexed by barcode, generating groups of reads originating from single cells. As a high-throughput and low-bias method of single-cell sequencing, SiC-seq will enable a broader range of genomic studies targeted at diverse cell populations. MyJove Corporation 2018-05-23 /pmc/articles/PMC6101372/ /pubmed/29889211 http://dx.doi.org/10.3791/57598 Text en Copyright © 2018, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Bioengineering
Demaree, Benjamin
Weisgerber, Daniel
Lan, Freeman
Abate, Adam R.
An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
title An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
title_full An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
title_fullStr An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
title_full_unstemmed An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
title_short An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
title_sort ultrahigh-throughput microfluidic platform for single-cell genome sequencing
topic Bioengineering
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6101372/
https://www.ncbi.nlm.nih.gov/pubmed/29889211
http://dx.doi.org/10.3791/57598
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