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BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing

Despite its widespread use, RNA-seq is still too laborious and expensive to replace RT-qPCR as the default gene expression analysis method. We present a novel approach, BRB-seq, which uses early multiplexing to produce 3′ cDNA libraries for dozens of samples, requiring just 2 hours of hands-on time....

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Autores principales: Alpern, Daniel, Gardeux, Vincent, Russeil, Julie, Mangeat, Bastien, Meireles-Filho, Antonio C. A., Breysse, Romane, Hacker, David, Deplancke, Bart
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6474054/
https://www.ncbi.nlm.nih.gov/pubmed/30999927
http://dx.doi.org/10.1186/s13059-019-1671-x
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author Alpern, Daniel
Gardeux, Vincent
Russeil, Julie
Mangeat, Bastien
Meireles-Filho, Antonio C. A.
Breysse, Romane
Hacker, David
Deplancke, Bart
author_facet Alpern, Daniel
Gardeux, Vincent
Russeil, Julie
Mangeat, Bastien
Meireles-Filho, Antonio C. A.
Breysse, Romane
Hacker, David
Deplancke, Bart
author_sort Alpern, Daniel
collection PubMed
description Despite its widespread use, RNA-seq is still too laborious and expensive to replace RT-qPCR as the default gene expression analysis method. We present a novel approach, BRB-seq, which uses early multiplexing to produce 3′ cDNA libraries for dozens of samples, requiring just 2 hours of hands-on time. BRB-seq has a comparable performance to the standard TruSeq approach while showing greater tolerance for lower RNA quality and being up to 25 times cheaper. We anticipate that BRB-seq will transform basic laboratory practice given its capacity to generate genome-wide transcriptomic data at a similar cost as profiling four genes using RT-qPCR. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13059-019-1671-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-64740542019-04-24 BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing Alpern, Daniel Gardeux, Vincent Russeil, Julie Mangeat, Bastien Meireles-Filho, Antonio C. A. Breysse, Romane Hacker, David Deplancke, Bart Genome Biol Method Despite its widespread use, RNA-seq is still too laborious and expensive to replace RT-qPCR as the default gene expression analysis method. We present a novel approach, BRB-seq, which uses early multiplexing to produce 3′ cDNA libraries for dozens of samples, requiring just 2 hours of hands-on time. BRB-seq has a comparable performance to the standard TruSeq approach while showing greater tolerance for lower RNA quality and being up to 25 times cheaper. We anticipate that BRB-seq will transform basic laboratory practice given its capacity to generate genome-wide transcriptomic data at a similar cost as profiling four genes using RT-qPCR. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13059-019-1671-x) contains supplementary material, which is available to authorized users. BioMed Central 2019-04-19 /pmc/articles/PMC6474054/ /pubmed/30999927 http://dx.doi.org/10.1186/s13059-019-1671-x Text en © The Author(s). 2019 Open AccessThis 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 Method
Alpern, Daniel
Gardeux, Vincent
Russeil, Julie
Mangeat, Bastien
Meireles-Filho, Antonio C. A.
Breysse, Romane
Hacker, David
Deplancke, Bart
BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing
title BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing
title_full BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing
title_fullStr BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing
title_full_unstemmed BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing
title_short BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing
title_sort brb-seq: ultra-affordable high-throughput transcriptomics enabled by bulk rna barcoding and sequencing
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6474054/
https://www.ncbi.nlm.nih.gov/pubmed/30999927
http://dx.doi.org/10.1186/s13059-019-1671-x
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