Benefits
Multiplexed full-length mRNA-seq
Full-length mRNA-seq library prep, 96 samples in a single tube.
Full-length transcript coverage
From differential gene expression to splicing variants and isoform detection.
Fast turnaround times
From samples to data in as little as one month.
What we do
MERCURIUS ™
Full-Length BRB-seq service
The MERCURIUS™ Full-Length BRB-seq service offers a convenient and streamlined solution for full-length mRNA transcriptomics projects of any size.
Clients can send us purified RNA samples, which are always quality-checked before launching our Full-Length BRB-seq pipeline. During the process, we always keep clients informed at defined checkpoints so that we can decide together how to best proceed to the next steps.
Next generation sequencing and data pre-processing (including alignment to the genome of choice) are part of our standard service as well. As a result, we provide our clients raw data, sequencing and alignment reports, and gene count matrices which can be used for downstream gene expression analysis.
Optional downstream differential gene expression analysis are also available upon request.
How does it work
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Client ships samples to Alithea
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1 week
Incoming QC - Client Checkpoint
(Nanodrop, Ribogreen, Tapestation) -
2 days
MERCURIUS™ Full-Length BRB-seq library preparation
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1 week
Library QC - Client Checkpoint
(Qubit, Fragment analyzer, shallow sequencing) -
1 week
Deep sequencing
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1 week
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Data
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Optional: Alithea performs Differential Gene Expression analysis
Performance
Uniform detection of 22'000+ genes at 12 million reads per sample across 48 samples
Distribution of the number of detected genes across 48...
Full-Length BRB-seq shows high demultiplexing rate and high mapping rate to exons
Mapping rates as high as 80% for different RNA inputs
Mapping rates are uniform across three RNA inputs...
Full-Length BRB-seq exhibits reads distribution across the transcript’s entire length
The gene body coverage shows a consistent and uniform...
>50'000 isoforms detected at 15M sequencing depth
Uniform detection of 22'000+ genes at 12 million reads per sample across 48 samples
Distribution of the number of detected genes across 48 samples prepared with the MERCURIUS™ Full-Length BRB-seq service. The library was sequenced at an average of 12 million reads.
Full-Length BRB-seq shows high demultiplexing rate and high mapping rate to exons
Mapping rates as high as 80% for different RNA inputs
Mapping rates are uniform across three RNA inputs tested: 1ug, 10ng, and 100ng. Human Universal Reference RNA was used to prepare mRNA Full-Length BRB libraries.
Full-Length BRB-seq exhibits reads distribution across the transcript’s entire length
The gene body coverage shows a consistent and uniform reads distribution across the entire gene body for the Full-Length BRB-seq protocol, comparable to the NEB protocol, while the BRB-seq protocol shows a significant 3' bias due to its poly-A selection methodology.
>50'000 isoforms detected at 15M sequencing depth
FAQs
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To generate high-quality sequencing data we recommend starting with 10 - 1000 ng of total purified RNA per sample.
The total RNA amount per pool should be at least 1000 ng.
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In addition to total RNA amount, it is important that the samples contain RNA of high integrity (RIN > 6) and are devoid of contaminants (Nanodrop A260/A230 between 1.8 and 2.2).
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Full-Length BRB-seq provides comprehensive coverage of the full-length mRNA transcripts.
We, therefore, normally recommend sequencing 12-20 million reads for each sample, which enables the reliable and unbiased detection of over 20,000 genes.
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As part of our standard service pipeline, we align the generated data to the genome of choice, provide a detailed report on the alignment and gene counting statistics and, finally, provide ready-to-use gene count matrices for downstream analysis.
Optionally, we can include differential gene expression analysis.
Speak with our RNA sequencing experts
Need guidance or have questions about BRB-seq and our service?
Easily book a call with our BRB-seq experts.