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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MyJove Corporation
2018
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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. |
format | Online Article Text |
id | pubmed-6101372 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
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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