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ddSeeker: a tool for processing Bio-Rad ddSEQ single cell RNA-seq data

BACKGROUND: New single-cell isolation technologies are facilitating studies on the transcriptomics of individual cells. Bio-Rad ddSEQ is a droplet-based microfluidic system that, when coupled with downstream Illumina library preparation and sequencing, enables the monitoring of thousands of genes pe...

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Autores principales: Romagnoli, Dario, Boccalini, Giulia, Bonechi, Martina, Biagioni, Chiara, Fassan, Paola, Bertorelli, Roberto, Sanctis, Veronica De, Leo, Angelo Di, Migliaccio, Ilenia, Malorni, Luca, Benelli, Matteo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6304778/
https://www.ncbi.nlm.nih.gov/pubmed/30583719
http://dx.doi.org/10.1186/s12864-018-5249-x
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author Romagnoli, Dario
Boccalini, Giulia
Bonechi, Martina
Biagioni, Chiara
Fassan, Paola
Bertorelli, Roberto
Sanctis, Veronica De
Leo, Angelo Di
Migliaccio, Ilenia
Malorni, Luca
Benelli, Matteo
author_facet Romagnoli, Dario
Boccalini, Giulia
Bonechi, Martina
Biagioni, Chiara
Fassan, Paola
Bertorelli, Roberto
Sanctis, Veronica De
Leo, Angelo Di
Migliaccio, Ilenia
Malorni, Luca
Benelli, Matteo
author_sort Romagnoli, Dario
collection PubMed
description BACKGROUND: New single-cell isolation technologies are facilitating studies on the transcriptomics of individual cells. Bio-Rad ddSEQ is a droplet-based microfluidic system that, when coupled with downstream Illumina library preparation and sequencing, enables the monitoring of thousands of genes per cell. Sequenced reads show unique features that do not permit the use of freely available tools to perform single cell demultiplexing. RESULTS: We present ddSeeker, a tool to perform initial processing and quality metrics of reads generated through Bio-Rad ddSEQ/Illumina experiments. Its application to the Illumina test dataset demonstrates that ddSeeker performs better than Illumina BaseSpace software, enabling a higher recovery of valid reads. We also show its utility in the analysis of an in-house dataset including two read sets characterized by low and high sequencing quality. ddSeeker and its source code are available at https://github.com/cgplab/ddSeeker. CONCLUSIONS: ddSeeker is a freely available tool to perform initial processing and quality metrics of reads generated through Bio-Rad ddSEQ/Illumina single cell transcriptomic experiments. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12864-018-5249-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-63047782019-01-02 ddSeeker: a tool for processing Bio-Rad ddSEQ single cell RNA-seq data Romagnoli, Dario Boccalini, Giulia Bonechi, Martina Biagioni, Chiara Fassan, Paola Bertorelli, Roberto Sanctis, Veronica De Leo, Angelo Di Migliaccio, Ilenia Malorni, Luca Benelli, Matteo BMC Genomics Software BACKGROUND: New single-cell isolation technologies are facilitating studies on the transcriptomics of individual cells. Bio-Rad ddSEQ is a droplet-based microfluidic system that, when coupled with downstream Illumina library preparation and sequencing, enables the monitoring of thousands of genes per cell. Sequenced reads show unique features that do not permit the use of freely available tools to perform single cell demultiplexing. RESULTS: We present ddSeeker, a tool to perform initial processing and quality metrics of reads generated through Bio-Rad ddSEQ/Illumina experiments. Its application to the Illumina test dataset demonstrates that ddSeeker performs better than Illumina BaseSpace software, enabling a higher recovery of valid reads. We also show its utility in the analysis of an in-house dataset including two read sets characterized by low and high sequencing quality. ddSeeker and its source code are available at https://github.com/cgplab/ddSeeker. CONCLUSIONS: ddSeeker is a freely available tool to perform initial processing and quality metrics of reads generated through Bio-Rad ddSEQ/Illumina single cell transcriptomic experiments. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12864-018-5249-x) contains supplementary material, which is available to authorized users. BioMed Central 2018-12-24 /pmc/articles/PMC6304778/ /pubmed/30583719 http://dx.doi.org/10.1186/s12864-018-5249-x Text en © The Author(s) 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Software
Romagnoli, Dario
Boccalini, Giulia
Bonechi, Martina
Biagioni, Chiara
Fassan, Paola
Bertorelli, Roberto
Sanctis, Veronica De
Leo, Angelo Di
Migliaccio, Ilenia
Malorni, Luca
Benelli, Matteo
ddSeeker: a tool for processing Bio-Rad ddSEQ single cell RNA-seq data
title ddSeeker: a tool for processing Bio-Rad ddSEQ single cell RNA-seq data
title_full ddSeeker: a tool for processing Bio-Rad ddSEQ single cell RNA-seq data
title_fullStr ddSeeker: a tool for processing Bio-Rad ddSEQ single cell RNA-seq data
title_full_unstemmed ddSeeker: a tool for processing Bio-Rad ddSEQ single cell RNA-seq data
title_short ddSeeker: a tool for processing Bio-Rad ddSEQ single cell RNA-seq data
title_sort ddseeker: a tool for processing bio-rad ddseq single cell rna-seq data
topic Software
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6304778/
https://www.ncbi.nlm.nih.gov/pubmed/30583719
http://dx.doi.org/10.1186/s12864-018-5249-x
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