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BRB-seq
Bulk RNA barcoding and sequencing (BRB-seq) is an ultra-high-throughput bulk 3' mRNA-seq technology that uses early-stage sample barcoding and unique molecular identifiers (UMIs) to allow the pooling of up to 384 samples in one tube early in the sequencing library preparation workflow. The transcriptomic technology is compatible with both Illumina and MGI short-read sequencing instruments.
In standard RNA-seq, a sequencing library must be prepared for each RNA sample individually. In contrast, in BRB-seq, all samples are pooled early in the workflow for simultaneous processing to reduce the cost and hands-on time associated with the library preparation stage
As BRB-seq is a 3' mRNA-seq technique, short reads are generated only for the 3' region of polyadenylated mRNA molecules instead of the full length of transcripts like in standard RNA-seq. This means that BRB-seq requires a far lower sequencing depth per sample to generate genome-wide transcriptomic data that allows users to detect similar numbers of expressed genes and differentially expressed genes as the standard Illumina TruSeq approach but at a cost up to 25 times cheaper or similar to profiling four genes using RT-qPCR. BRB-seq also has a greater tolerance for lower RNA quality (RIN <6) where transcripts are degraded because only the 3' region is required in library preparation
The BRB-seq technique was first published in April 2019 in the peer-reviewed journal Genome Research in a manuscript entitled 'BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing. By the end of 2019, the article was among the top 10 most-read papers in the journal and has been cited over 150 times (April 2024).
The technique was developed at the École Polytechnique Fédérale de Lausanne in Switzerland in the labs of Professor Bart Deplancke and collaborators. In May 2020, a company called Alithea Genomics was established to provide BRB-seq as kits for researchers or as a full service. BRB-seq builds upon technological advances in single-cell transcriptomics, where sample barcoding made the early multiplexing of hundreds to thousands of single cells possible. Sample multiplexing allowed researchers to create single sequencing libraries containing multiple distinct samples, reducing overall experimental costs and hands-on time while dramatically boosting throughput.
BRB-seq applies these advancements in sample and mRNA barcoding to mRNAs derived from bulk cell populations to enable ultra-high-throughput studies crucial for drug discovery, population studies, or fundamental research.
The fundamental aspect of BRB-seq is the optimized sample barcode primers. Each barcoded nucleotide sequence includes an adaptor for primer annealing, a 14-nt long barcode that assigns a unique identifier to each individual RNA sample, and a random 14-nt long UMI that tags each mRNA molecule with a unique sequence to distinguish between original mRNA transcripts and duplicates that result from PCR amplification bias. BRB-seq allows up to 384 individually barcoded RNA samples to be pooled into one tube early in the workflow to streamline subsequent steps in cDNA library preparation and sequencing.
Input RNA requirements
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BRB-seq AI simulator
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BRB-seq
Bulk RNA barcoding and sequencing (BRB-seq) is an ultra-high-throughput bulk 3' mRNA-seq technology that uses early-stage sample barcoding and unique molecular identifiers (UMIs) to allow the pooling of up to 384 samples in one tube early in the sequencing library preparation workflow. The transcriptomic technology is compatible with both Illumina and MGI short-read sequencing instruments.
In standard RNA-seq, a sequencing library must be prepared for each RNA sample individually. In contrast, in BRB-seq, all samples are pooled early in the workflow for simultaneous processing to reduce the cost and hands-on time associated with the library preparation stage
As BRB-seq is a 3' mRNA-seq technique, short reads are generated only for the 3' region of polyadenylated mRNA molecules instead of the full length of transcripts like in standard RNA-seq. This means that BRB-seq requires a far lower sequencing depth per sample to generate genome-wide transcriptomic data that allows users to detect similar numbers of expressed genes and differentially expressed genes as the standard Illumina TruSeq approach but at a cost up to 25 times cheaper or similar to profiling four genes using RT-qPCR. BRB-seq also has a greater tolerance for lower RNA quality (RIN <6) where transcripts are degraded because only the 3' region is required in library preparation
The BRB-seq technique was first published in April 2019 in the peer-reviewed journal Genome Research in a manuscript entitled 'BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing. By the end of 2019, the article was among the top 10 most-read papers in the journal and has been cited over 150 times (April 2024).
The technique was developed at the École Polytechnique Fédérale de Lausanne in Switzerland in the labs of Professor Bart Deplancke and collaborators. In May 2020, a company called Alithea Genomics was established to provide BRB-seq as kits for researchers or as a full service. BRB-seq builds upon technological advances in single-cell transcriptomics, where sample barcoding made the early multiplexing of hundreds to thousands of single cells possible. Sample multiplexing allowed researchers to create single sequencing libraries containing multiple distinct samples, reducing overall experimental costs and hands-on time while dramatically boosting throughput.
BRB-seq applies these advancements in sample and mRNA barcoding to mRNAs derived from bulk cell populations to enable ultra-high-throughput studies crucial for drug discovery, population studies, or fundamental research.
The fundamental aspect of BRB-seq is the optimized sample barcode primers. Each barcoded nucleotide sequence includes an adaptor for primer annealing, a 14-nt long barcode that assigns a unique identifier to each individual RNA sample, and a random 14-nt long UMI that tags each mRNA molecule with a unique sequence to distinguish between original mRNA transcripts and duplicates that result from PCR amplification bias. BRB-seq allows up to 384 individually barcoded RNA samples to be pooled into one tube early in the workflow to streamline subsequent steps in cDNA library preparation and sequencing.
Input RNA requirements