Bulk RNA sequencing High-quality transcriptome data
Our Bulk RNA sequencing is based on single-cell chemistry, enabling low-input sequencing. Bulk RNA sequencing is optimal for sequencing intact polyadenylated RNA, whereas Total Bulk RNA sequencing allows for the analysis of full-length RNA and a diverse range of RNA species.
Features of bulk RNA sequencing
Detect transcriptome profiles in populations
Bulk RNA sequencing (Bulk-seq) is the method of choice for transcriptomic analysis of pooled cell populations, tissue sections, and biopsies. It measures the average expression level of individual genes across hundreds to millions of input cells to produce a global idea of gene expression differences between samples.
We use the same CEL-Seq2–style barcodes and reagents as for our SORT-seq technology to enable sample multiplexing. Each sample gets its own sample-specific barcode, which allows us to pool multiple samples into one sequencing library. Our use of single-cell sequencing reagents allows us to process bulk samples with as few as 100 cells as input. For high-throughput RNA sequencing, we recommend to make use of our Discovery-seq service.
The ability to process even very low–input samples can sometimes make the difference when you are investigating rare cell types, are interested in select populations, or require a complex experimental design with few surviving cells. We have performed bulk RNA-seq on more than twenty different species and we are experienced with various tissue types, from cultured dendritic cells to transgenic brain tumors and from intestinal organoids to microalgae.
Total Bulk RNA Sequencing
Total Bulk RNA sequencing
The Total Bulk RNA sequencing (Total Bulk-seq) service leverages the familiarity of Bulk RNA sample preparation, including options for both normal and low input, and the precision of Cel-seq2 barcoding. This service combines these techniques with full-length total RNA sequencing, allowing for comprehensive analysis.
Unlike traditional Bulk-seq, which reads the 3’ ends of polyadenylated RNA, our Total Bulk RNA sequencing provides full-length RNA sequencing, ensuring a more complete and detailed transcriptomic profile.
A standard protocol for Total Bulk RNA sequencing is available for human, mouse, and rat samples. Are you working with other species? Do not hesitate to contact us to see what we can do for you.
Bulk RNA sequencing
Information Guide
Get an overview of Single Cell Discoveries, Bulk RNA sequencing, how to get started, and more in our Bulk RNA sequencing information guide.
Benefits
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Directly comparable with SORT-seq data
Our bulk RNA sequencing service is based on the same chemistry as our SORT-seq technology for single-cell sequencing. This means that you can directly compare the results from your single-cell sequencing experiments with the results from your bulk RNA analysis. -
Available for low-input
Because we use single-cell reagents, we can perform a bulk RNA sequencing analysis on extremely low sample volumes. We need only 100 cells for bulk RNA sequencing. -
Combine multiple samples in one sequencing library
Our ability to multiplex samples means we can speed up turnaround times and enhance the processing efficiency of large amounts of samples.
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Directly comparable with VASA-seq data
Analysis of full-length transcriptomes is possible. This enables you to study alternative splicing and make RNA velocity predictions. You can directly compare results from your single-cell VASA-seq experiments with your Total Bulk RNA analysis results.
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Available for low-input
Because we use single-cell reagents, we can perform a Total Bulk RNA sequencing analysis on extremely low sample volumes. We need only 100 cells for Total Bulk RNA sequencing.
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Rare cell type processing
We accept cell pellets and FFPE samples for Total Bulk RNA sequencing, providing flexibility in sample collection and preparation, and making the process less time-sensitive. This is especially useful when working with rare cell types. We are also able to sequence microorganisms.
Step 1
Consultation
Every project starts with one or more free consultations because no two single-cell projects are the same. Our experienced PhD-level team discusses your biological question, sample type, planning, data analysis options, and other wishes. This will ensure you select the best method for your project and can hit the ground running.
Step 2
Sample Preparation
You can decide to perform RNA purification yourself or outsource it to our laboratory. Check the sample requirements for details on both options.
Step 3
Processing in Our Lab
Our experienced team performs the bulk RNA sequencing protocol in our lab. We sequence the libraries on our Illumina NovaSeq X Plus or NextSeq 2000. We'll do multiple QC checks and keep you informed of the status of your project.
Step 4
Data Analysis
Our data team performs data quality checks and discusses irregularities with our lab team if necessary. You will receive the raw data files, data report, and a guiding explanation. Talk to our data consulting team for help or request additional investigations such as gene set enrichment analysis and deconvolution of cell type abundances.
Deliverables
As with all of our services, we provide ongoing support from our single-cell sequencing specialists. For each bulk RNA sequencing project, we provide the following.
- Regular updates during sample processing, including QC
- Raw sequencing data (FASTQ files)
- Gene count tables
- QC report of mapping results (raw and mapped reads per sample, and PCA plot)
- An explanation from our data team about data quality metrics
- All raw sequencing data, results and reports are processed on EU servers, and securely encrypted using an AES (Advanced Encryption Standard) algorithm with a 256-bit key, before being delivered to the customers and archived.
CRISPR screening at scale
Achieving 1 million cell data sets with an instrument-free, cost-effective approach.
Recent publications using bulk RNA sequencing
Science Advances
Isolation and tracing of matrix-producing notochordal and chondrocyte cells using ACAN-2A-mScarlet reporter human iPSC lines
Cell Reports Medicine
Myeloid cells coordinately induce glioma cell-intrinsic and cell-extrinsic pathways for chemoresistance via GP130 signaling
FAQ
All questions below apply to Bulk RNA sequencing and Total Bulk RNA sequencing.
Sequencing depth depends on the number of samples, your budget, and your biological question. We offer two packages for Bulk RNA sequencing: STANDARD, with 10 million sequencing reads per sample, and PLUS, with 20 million sequencing reads per sample.
Even Bulk RNA sequencing is not without noise. Hence it is usually wise for downstream analysis purposes to consider triplicates or, at least, duplicates in your experimental set-up. We always recommend you include at least a second vial for each of the samples to ensure technical backup.
We perform STAR software for data mapping and DEseq2 for differential gene expression analysis.
Pricing depends on the cells per sample, sequencing depth, viability of input materials, and required protocol modifications. Your account manager can give pricing information specific to your project’s requirements during consultations.
Information Guide
Discover our bulk RNA sequencing service
Download our information guide to access an overview of Single Cell Discoveries, explore our bulk RNA sequencing service, learn how to get started, and gain more valuable insights.