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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....
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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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. |
format | Online Article Text |
id | pubmed-6474054 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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