Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Bulk RNA Barcoding and sequencing (BRB-seq) protocol for RNA-seq of human pre-adipocytes and differentiated adipocytes


ABSTRACT: Here, we validate a novel protocol, Bulk RNA Barcoding and sequencing (BRB-seq), that combines the multiplexing-driven cost-effectiveness of a single-cell RNA-seq protocol with the efficiency of a bulk RNA-seq procedure. For this we use BRB-seq protocol on human pre-adipocytes and differentiated adipocytes, and compare its effectiveness as compared to TruSeq. Indeed, one of the principal limitations of bulk RNA-seq is the time and costs of library preparation, which makes it difficult to profile many samples simultaneously. Here, BRB-seq produces 3’ libraries that exhibit similar gene expression quantification to TruSeq and maintain this quality even with low quality RNA samples.

INSTRUMENT(S): NextSeq 500

ORGANISM(S): Homo sapiens

SUBMITTER: Vincent Gardeux 

PROVIDER: E-MTAB-6469 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

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

Alpern Daniel D   Gardeux Vincent V   Russeil Julie J   Mangeat Bastien B   Meireles-Filho Antonio C A ACA   Breysse Romane R   Hacker David D   Deplancke Bart B  

Genome biology 20190419 1


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  ...[more]

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