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Single-cell isoform analysis in human immune cells

High-throughput single-cell analysis today is facilitated by protocols like the 10X Genomics platform or Drop-Seq which generate cDNA pools in which the origin of a transcript is encoded at its 5′ or 3′ end. Here, we used R2C2 to sequence and demultiplex 12 million full-length cDNA molecules generat...

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Autores principales: Volden, Roger, Vollmers, Christopher
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8819920/
https://www.ncbi.nlm.nih.gov/pubmed/35130954
http://dx.doi.org/10.1186/s13059-022-02615-z
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author Volden, Roger
Vollmers, Christopher
author_facet Volden, Roger
Vollmers, Christopher
author_sort Volden, Roger
collection PubMed
description High-throughput single-cell analysis today is facilitated by protocols like the 10X Genomics platform or Drop-Seq which generate cDNA pools in which the origin of a transcript is encoded at its 5′ or 3′ end. Here, we used R2C2 to sequence and demultiplex 12 million full-length cDNA molecules generated by the 10X Genomics platform from ~3000 peripheral blood mononuclear cells. We use these reads, independent from Illumina data, to identify B cell, T cell, and monocyte clusters and generate isoform-level transcriptomes for cells and cell types. Finally, we extract paired adaptive immune receptor sequences unique to each T and B cell. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13059-022-02615-z.
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spelling pubmed-88199202022-02-08 Single-cell isoform analysis in human immune cells Volden, Roger Vollmers, Christopher Genome Biol Method High-throughput single-cell analysis today is facilitated by protocols like the 10X Genomics platform or Drop-Seq which generate cDNA pools in which the origin of a transcript is encoded at its 5′ or 3′ end. Here, we used R2C2 to sequence and demultiplex 12 million full-length cDNA molecules generated by the 10X Genomics platform from ~3000 peripheral blood mononuclear cells. We use these reads, independent from Illumina data, to identify B cell, T cell, and monocyte clusters and generate isoform-level transcriptomes for cells and cell types. Finally, we extract paired adaptive immune receptor sequences unique to each T and B cell. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13059-022-02615-z. BioMed Central 2022-02-07 /pmc/articles/PMC8819920/ /pubmed/35130954 http://dx.doi.org/10.1186/s13059-022-02615-z Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Method
Volden, Roger
Vollmers, Christopher
Single-cell isoform analysis in human immune cells
title Single-cell isoform analysis in human immune cells
title_full Single-cell isoform analysis in human immune cells
title_fullStr Single-cell isoform analysis in human immune cells
title_full_unstemmed Single-cell isoform analysis in human immune cells
title_short Single-cell isoform analysis in human immune cells
title_sort single-cell isoform analysis in human immune cells
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8819920/
https://www.ncbi.nlm.nih.gov/pubmed/35130954
http://dx.doi.org/10.1186/s13059-022-02615-z
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