The MERCURIUS™ Blood BRB-seq library preparation kits for Illumina® contain all the oligos and enzymes needed to go from purified blood RNA to sequencing-ready DNA libraries.
Bulk RNA sequencing at scale
More samples, more replicates. Robust results, significant discoveries.
Streamlined data pre-processing
Demultiplex and align your BRB-seq data with our easy-to-use cloud-based platform.
Integrated globin depletion
Seamlessly integrated globin depletion. No need to purchase additional kits.
One-day lab workflow
Convenient and short protocol from samples to sequencing-ready libraries in one day.
The generated 3' mRNA-seq data is ideally suited for differential gene expression analysis.
As compared to other bulk 3' mRNA-seq solutions, BRB-seq makes it possible to process virtually any number of blood RNA samples in one single tube, which helps reduce technical variation, turnaround times and costs.
The particular aspect of the Blood BRB-seq workflow is the addition of proprietary globin blockers directly at the reverse transcription stage, which in turns enable efficient and effortless depletion of unwanted globin genes.
Each kit contains enough reagents (including Unique Dual Indexing oligos) to prepare up-to four sequencing libraries.
After sequencing, the generated data can be easily demultiplexed and aligned to the genome of choice using our dedicated cloud-based pipeline, which does not require prior bioinformatic experience.
Experimental workflow at a glance
1. Reverse Transcription
2. Samples Pooling
3. Second strand synthesis
5. Library indexing and amplification
6. Library QC and sequencing
7. Sequencing data processing and analysis
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.
The recommended range of RNA amount for each sample is of 50ng-1μg, normally the more RNA, the better.
The minimum recommended RIN number is 6 and the A260/230 ratio (Nanodrop) should be in the 1.5-2.2 range.
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. In order to facilate as much as possible BRB-seq data analysis, we have also created a dedicate pipeline which can be found in our “SOFTWARE” page.
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.
Speak with our RNA sequencing experts
Book a one-on-one call with one of our RNA experts to discover how we can assist your next project.