BRB-seq technology

MERCURIUS™ BRB-seq is a transformative tool combining unbiased, ultra-high-content, and high-throughput screening with massively parallel transcriptomics.

This method uses highly optimized and rigorously evaluated sample barcodes and unique molecular identifiers to tag the 3’ poly(A) tail of all mRNA molecules in a sample-specific manner during the first-strand synthesis step of cDNA library preparation. After this step, all samples can be pooled into one single tube and processed together for the remainder of the library prep workflow. 

The BRB-seq protocol is designed to work with purified RNA samples from all eukaryotic species. 

MERCURIUS™ BRB-seq stands out from traditional RNA-seq methods by offering a high-throughput, cost-efficient, and streamlined workflow specifically designed for large-scale screening applications. One of the key differences lies in its sample multiplexing strategy: BRB-seq allows multiple RNA samples to be barcoded and pooled at the earliest step of the protocol—right after the RT reaction—so that the entire library preparation can proceed in a single tube. This significantly reduces both hands-on time and reagent costs.

In contrast, standard RNA-seq workflows typically require individual processing of all the RNA samples. This makes them more labor-intensive, costly, and less scalable, especially when working with large numbers of conditions, compounds, or replicates—common in drug discovery pipelines.

Each BRB-seq kit contains reagents (including four pairs of Unique Dual Indexing adapters) sufficient for the complete library preparation process for four different BRB-seq pools. To note, the total number of RNA samples that can be processed with one kit does not exceed the kit specifications; for instance, a 96-samples kit can be used to prepare up-to 96 samples distributed across up-to four different libraries.

One of the key advantages of BRB-seq is that it does not only save reagents and cost in the library preparation stage, but also in the sequencing one. As opposed to standard RNA-seq, where 20M-30M reads per sample are required, we normally recommend to sequence BRB-seq libraries at a depth of 4M-5M reads per sample, which is normally enough to detect the vast majority of expressed genes.

The only difference between BRB-seq and standard RNA-seq data analysis is the demultiplexing step, which is used to assign sequencing reads to their sample of origin based on the BRB-seq barcode sequence. For a thorough description of BRB-seq data processing, please refer to the BRB-seq kit user guide.