scPipe: A flexible R/Bioconductor preprocessing pipeline for single-cell RNA-sequencing data.

Single-cell RNA sequencing (scRNA-seq) technology allows researchers to profile the transcriptomes of thousands of cells simultaneously. Protocols that incorporate both designed and random barcodes have greatly increased the throughput of scRNA-seq, but give rise to a more complex data structure. Th...

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Main Authors: Luyi Tian, Shian Su, Xueyi Dong, Daniela Amann-Zalcenstein, Christine Biben, Azadeh Seidi, Douglas J Hilton, Shalin H Naik, Matthew E Ritchie
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2018-08-01
Series:PLoS Computational Biology
Online Access:https://doi.org/10.1371/journal.pcbi.1006361
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spelling doaj-816479d2bf604f45a983e4b4c954ccad2021-04-21T15:12:40ZengPublic Library of Science (PLoS)PLoS Computational Biology1553-734X1553-73582018-08-01148e100636110.1371/journal.pcbi.1006361scPipe: A flexible R/Bioconductor preprocessing pipeline for single-cell RNA-sequencing data.Luyi TianShian SuXueyi DongDaniela Amann-ZalcensteinChristine BibenAzadeh SeidiDouglas J HiltonShalin H NaikMatthew E RitchieSingle-cell RNA sequencing (scRNA-seq) technology allows researchers to profile the transcriptomes of thousands of cells simultaneously. Protocols that incorporate both designed and random barcodes have greatly increased the throughput of scRNA-seq, but give rise to a more complex data structure. There is a need for new tools that can handle the various barcoding strategies used by different protocols and exploit this information for quality assessment at the sample-level and provide effective visualization of these results in preparation for higher-level analyses. To this end, we developed scPipe, an R/Bioconductor package that integrates barcode demultiplexing, read alignment, UMI-aware gene-level quantification and quality control of raw sequencing data generated by multiple protocols that include CEL-seq, MARS-seq, Chromium 10X, Drop-seq and Smart-seq. scPipe produces a count matrix that is essential for downstream analysis along with an HTML report that summarises data quality. These results can be used as input for downstream analyses including normalization, visualization and statistical testing. scPipe performs this processing in a few simple R commands, promoting reproducible analysis of single-cell data that is compatible with the emerging suite of open-source scRNA-seq analysis tools available in R/Bioconductor and beyond. The scPipe R package is available for download from https://www.bioconductor.org/packages/scPipe.https://doi.org/10.1371/journal.pcbi.1006361
collection DOAJ
language English
format Article
sources DOAJ
author Luyi Tian
Shian Su
Xueyi Dong
Daniela Amann-Zalcenstein
Christine Biben
Azadeh Seidi
Douglas J Hilton
Shalin H Naik
Matthew E Ritchie
spellingShingle Luyi Tian
Shian Su
Xueyi Dong
Daniela Amann-Zalcenstein
Christine Biben
Azadeh Seidi
Douglas J Hilton
Shalin H Naik
Matthew E Ritchie
scPipe: A flexible R/Bioconductor preprocessing pipeline for single-cell RNA-sequencing data.
PLoS Computational Biology
author_facet Luyi Tian
Shian Su
Xueyi Dong
Daniela Amann-Zalcenstein
Christine Biben
Azadeh Seidi
Douglas J Hilton
Shalin H Naik
Matthew E Ritchie
author_sort Luyi Tian
title scPipe: A flexible R/Bioconductor preprocessing pipeline for single-cell RNA-sequencing data.
title_short scPipe: A flexible R/Bioconductor preprocessing pipeline for single-cell RNA-sequencing data.
title_full scPipe: A flexible R/Bioconductor preprocessing pipeline for single-cell RNA-sequencing data.
title_fullStr scPipe: A flexible R/Bioconductor preprocessing pipeline for single-cell RNA-sequencing data.
title_full_unstemmed scPipe: A flexible R/Bioconductor preprocessing pipeline for single-cell RNA-sequencing data.
title_sort scpipe: a flexible r/bioconductor preprocessing pipeline for single-cell rna-sequencing data.
publisher Public Library of Science (PLoS)
series PLoS Computational Biology
issn 1553-734X
1553-7358
publishDate 2018-08-01
description Single-cell RNA sequencing (scRNA-seq) technology allows researchers to profile the transcriptomes of thousands of cells simultaneously. Protocols that incorporate both designed and random barcodes have greatly increased the throughput of scRNA-seq, but give rise to a more complex data structure. There is a need for new tools that can handle the various barcoding strategies used by different protocols and exploit this information for quality assessment at the sample-level and provide effective visualization of these results in preparation for higher-level analyses. To this end, we developed scPipe, an R/Bioconductor package that integrates barcode demultiplexing, read alignment, UMI-aware gene-level quantification and quality control of raw sequencing data generated by multiple protocols that include CEL-seq, MARS-seq, Chromium 10X, Drop-seq and Smart-seq. scPipe produces a count matrix that is essential for downstream analysis along with an HTML report that summarises data quality. These results can be used as input for downstream analyses including normalization, visualization and statistical testing. scPipe performs this processing in a few simple R commands, promoting reproducible analysis of single-cell data that is compatible with the emerging suite of open-source scRNA-seq analysis tools available in R/Bioconductor and beyond. The scPipe R package is available for download from https://www.bioconductor.org/packages/scPipe.
url https://doi.org/10.1371/journal.pcbi.1006361
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