MERCURIUS™
Total BRB-seq Kits
for multipliplexed full-length total RNA-seq
MERCURIUS™ Total BRB-seq combines rRNA depletion, full-length total RNA sequencing, and early sample multiplexing for scalable whole-transcriptome profiling from purified RNA.
Multiplexed
full-length RNA
Full-length transcript coverage
For a variety of samples and species
One-day library prep workflow
Product Overview
MERCURIUS™ Total BRB-seq is a total RNA-seq kit for purified RNA samples, designed for scalable whole-transcriptome profiling when coding and non-coding transcript information is required.
The workflow combines rRNA depletion, early sample barcoding, and multiplexing of up to 96 purified RNA samples in a single tube. By preserving full-length transcript coverage, MERCURIUS™ Total BRB-seq supports applications such as differential expression analysis, regulatory RNA studies, isoform and splice variant analysis, biomarker discovery, toxicogenomics, and richer transcriptomic dataset generation.
at 10M reads per sample
Why use total RNA-seq instead of mRNA-seq?
Total RNA-seq enables broader transcriptome profiling than poly(A)-selected or 3′ mRNA-seq workflows by capturing both coding and non-coding RNA species. This makes it useful when regulatory RNAs, antisense transcripts, isoforms, splice variants, or broader whole-transcriptome information may contribute to the biological question.
For gene-level expression profiling at the lowest cost per sample, 3′ BRB-seq may be the better fit. For broader transcript coverage, choose Total BRB-seq.
Applications for total RNA-seq from purified RNA
- Whole-transcriptome profiling
Capture coding and non-coding transcript information from purified RNA samples. - Differential expression analysis
Compare gene expression across conditions, treatments, genotypes, disease states, or time points. - Regulatory RNA studies
Profile long non-coding RNAs, antisense transcripts, and other RNA classes that may be missed by mRNA-focused workflows. - Isoform and splice variant analysis
Use full-length transcript coverage when 3′ mRNA-seq is not sufficient. - Toxicogenomics and pathway analysis
Study broader transcriptomic responses to compounds, exposures, or biological perturbations. - Biomarker discovery and translational research
Build richer RNA-seq datasets from purified RNA for disease, response, or phenotype-associated signatures.
Total BRB-seq Matches NEBNext® Ultra™ II mRNA and Total RNA Gene Detection Sensitivity
Saturation curves indicating the number of detected genes as a function of sequencing depth in Huh7 cells. MERCURIUS™ Full-Length BRB-seq and Total BRB-seq perform comparably to NEBNext® Ultra™ II mRNA and Total RNA preparations, respectively, demonstrating equivalent sensitivity at a fraction of the library preparation cost and effort. MERCURIUS™ BRB-seq is a 3’ mRNA library preparation technology, hence the lower overall gene detection versus full-length methods.
Total BRB-seq Matches NEBNext® Ultra™ II mRNA and Total RNA Transcript Detection Sensitivity
Saturation curves indicating the number of detected transcripts as a function of sequencing depth in Huh7 cells. MERCURIUS™ Full-Length BRB-seq and Total BRB-seq perform comparably to NEBNext® Ultra™ II mRNA and Total RNA preparations, respectively, demonstrating equivalent sensitivity at a fraction of the library preparation cost and effort. MERCURIUS™ BRB-seq specifically detects protein-coding transcripts, hence the lower overall transcript detection compared to full-length methods.
4. Total BRB-seq Clusters Tightly with its NEBNext® Ultra™ II Counterpart and Resolves TGF-β Treatment Effects
Principal component analysis (PCA) of gene expression profiles generated by the specified library preparation methods across untreated (circles) and TGF-β-treated (triangles) Huh7 samples. Samples cluster primarily by library type along PC2 and by treatment condition along PC1. All methods robustly capture the expression response to treatment. MERCURIUS™ Full-Length BRB-seq and Total BRB-seq cluster closely with their NEBNext® Ultra™ II counterparts, demonstrating equivalent transcriptomic profiles. MERCURIUS™ BRB-seq clusters separately, consistent with its 3′-end capture design.
5. Total BRB-seq Shares the Majority of Differentially Expressed Genes and Transcripts with its NEBNext® Ultra™ II Counterpart
Venn diagrams showing the overlap in differentially expressed (DE) genes (left) and transcripts (right) identified by MERCURIUS™ Full Length BRB-seq and NEBNext® Ultra™ II for mRNA libraries (top) and total RNA libraries (bottom). Numbers indicate DE features unique to each method or shared between both. At the gene level, MERCURIUS™ Full Length BRB-seq and NEBNext® Ultra™ II share over 88% of DE genes for mRNA and 84% for total RNA preparations, demonstrating strong concordance in differential expression calls across both library types. The overlap is smaller but still strong for DE transcripts across technologies.
6. Total BRB-seq Reproduces Differential Expression Results with Excellent Concordance to its NEBNext® Ultra™ II Counterpart
Log2 fold-change correlation between MERCURIUS™ Full-Length BRB-seq and NEBNext® Ultra™ II for mRNA (left, R = 0.983) and total RNA (right, R = 0.980) protocols, across intersecting gene sets of 13,364 and 13,237 genes, respectively. Each point represents a gene, colored by its differential expression (DE) or non-significant (NS) status after treatment of Huh7 cells with human TGF-β versus untreated cells. The dashed diagonal represents perfect correlation. The concordance of DE fold changes across both RNA input types demonstrates that MERCURIUS™ Full-Length and Total BRB-seq reliably recapitulate the results of gold-standard library preparation.
Product documentation
Publications
- 3' BRB-seq, BRB-seq
Larissa Carvalho Costa, Christopher Gottschalk, Davis M. Upchurch, Oscar P. Hurtado-Gonzales, Ben N. Mansfeld, Alan Yocca, Lauren Whitt, Cheryl Vann, Gennaro Fazio
- 3' BRB-seq, BRB-seq
Stéphanie Boder‐Pasche, Mustafa Demir, Sarah Heub, Manon Garzuel, Réal Ischer, Daniel Migliozzi, Siegfried Graf, Noa Schmid, H. Baris Atakan, Daria Gudkova, Daniel Alpern, Riccardo Dainese, Bart Deplancke, Gilles Weder
- 3' BRB-seq, BRB-seq
Maxine Leonardi, Yves Paychère, Felix Naef.
Application
Equipment compatibility
Species compatibility
Available formats
Shipping conditions
Storage conditions
What is a total RNA-seq kit?
A total RNA-seq kit prepares sequencing libraries from total RNA, enabling broader transcriptome profiling than mRNA-only approaches. Depending on the workflow, total RNA-seq can capture coding and non-coding transcripts after ribosomal RNA depletion.
What is the maximum number of pools I can prepare with one kit?
Each kit includes 4 Unique Dual Index (UDI) pairs, so you can prepare up to 4 indexed libraries (pools) per kit.
What’s the RNA input range and quality requirement?
The tested input range is 100 pg to 100 ng total RNA per well. For best results (library complexity + uniformity), aim for 10–100 ng per well, and pool at least 8 wells per library.
Because Total BRB-seq uses early multiplexing, it’s important that RNA quantity and quality are consistent across samples (e.g., RIN > 7, 260/230 > 1.5, and ~±10% input uniformity).
What does “Total” mean in Total BRB-seq?
Total BRB-seq is designed for total RNA sequencing (not just mRNA), including coding and non-coding transcripts, and includes an rRNA depletion step prior to library prep.
What sequencing depth should I plan for?
We recommend 10-20 million reads for each sample, which enables the detection of 65,000 transcripts.
Does bioinformatic processing differ from standard bulk RNA-seq?
Yes, there is one extra step: after standard index demultiplexing, you demultiplex by sample barcodes to generate per-sample gene/transcript count outputs. The workflow supports producing outputs like UMI count matrices, gene count matrices, and a QC report.
Where do I find the barcode sequences for demultiplexing?
Go to the “Resources” section of this page and click on the button to download the barcode files for the product you are using.
Can I run Total BRB-seq as a service instead of in-house?
Yes. If you are not ready to run kits internally, Alithea can run the workflow and deliver FASTQ files, gene count matrices, and analysis report files.
Ask your question:
Kit name
Catalog number
preps
pairs
MERCURIUS™ Total BRB-seq library prep kit (96 preps)
all eukaryotic species*
MERCURIUS™ Total BRB-seq library prep kit (4x96 preps)
all eukaryotic species*
* Contact us to inquire about compatibility with other species.
Accessories
Product
Catalog Number
MERCURIUS™ X-Leap-compatible MF.UDI Expansion module
MERCURIUS™ X-Leap-compatible MF.UDI Expansion module
MERCURIUS™ Full-Length Post-Pooling Preparation Module (4 libraries)
Ordering tip
Each kit includes 4 UDI pairs. Add the expansion module if you need more unique indexes (total 16 UDI pairs available).
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