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Omics Research // Transcriptomics

Total RNA Sequencing

Accurately measure the number of genes and transcripts in all coding and non-coding RNA molecules within the transcriptome. Unlock insight into disease states with a variety of sequencing strategies for mRNA, tRNA, lncRNA, and sncRNA species.

What Is Total RNA Sequencing?

The next-generation sequencing process of sequencing mRNA with the inclusion of non-coding RNA (ncRNA).


Why is Total RNA Sequencing Important?

RNA transcription is the link between DNA and protein expression.

Studying the RNA present gives scientists insight into the biology of a cell and how changes — alternative splicing events, gene fusions, and allele-specific expression of rare and novel messages — may relate to disease.

What Can You Do?

Ribosomal RNA Depletion

Remove abundant rRNA molecules to focus on the coding and non-coding transcripts of interest


Strand-Specific Sequencing

Identify which strand of DNA the transcript was derived from


Low-Input RNA Sequencing

Use less than 10 ng of extracted total RNA. Learn more about low-input seq


Experienced Bioinformatics Experts

Basic gene expression and SNV/InDel or advanced bioinformatic analysis services

Technologies Used for Total RNA Sequencing

rRNA Depletion

Total RNA sequencing uses a ribosomal RNA (rRNA) depletion step to remove the majority of rRNA before preparing libraries of your RNA. This negative selection approach enables the researcher to increase the amount of relevant sequencing reads and examine:

  • messenger RNA (mRNA)
  • long non-coding RNA (lncRNA)
  • small non-coding RNA (sncRNA)
  • microRNA (miRNA)
  • small nuclear RNA (snoRNA)
  • piwi-interacting RNA (piRNA)
  • tRNA derived small RNA (tsRNA)

Stranded RNAseq

Psomagen uses strand-specific total RNA sequencing, or stranded RNAseq, to identify overlapping transcripts and genes.

Researchers can use the strand orientation information in their analyses to aid in annotation and novel transcript calls.

Low-Input Stranded RNAseq

Don’t have a lot of RNA? Use our low-input stranded RNAseq service. This service still provides high quality data for both coding and non-coding RNA by enabling high sensitivity through changing the order in which the ribosomal RNA signal is targeted. The ribosomal RNA depletion step is skipped at the beginning of the workflow and alternatively, ribosomal cDNA depletion occurs after library assembly.

It is possible to use our low-input stranded RNAseq service with RNA input as low as 250pg.

On-Going Promotions

We're always adding new deals — check in often for updated promotions!


Total RNA Sequencing Best Practices

Sample Preparation & RNA Extraction

Total RNA extraction from cells, tissue, FFPE, blood, and swab samples

  • Sample QC: >500 ng and RIN >7.0
  • FFPE: >200 ng and DV 200 >70%

For blood samples, globin mRNA depletion can be performed to reduce its abundance and investigate other mRNAs.

Have a low amount of samples? Take a look at our low-input RNA seq to find the right option for you.

Library Preparation

Stranded Total RNA

  • 500 ng Total RNA (100-1000 ng)

Low-Input Total RNA

  • 10 ng Total RNA (250 pg-10 ng)

Sequencing Instrumentation & Read Length

Illumina NovaSeq 6000

  • 2 x 150 bp read length
  • 200-240 samples/lane (20M reads)

To assess novel alternative transcripts and RNA editing, we recommend a targeted read depth of at least 50 million

Psomagen can only guarantee raw sequencing depth

Bioinformatic Analysis

Basic analysis:

  • Mapping
  • Gene expression profiling
  • Variant analysis (SNV, InDels)

Advanced analysis:

  • Novel alternative transcripts
  • Fusion gene list
  • Differentially Expressed Gene (DEG)
  • KEGG Enrichment Analysis
  • Alternatively spliced transcripts

Data Delivery

All sequencing run data is delivered as demultiplexed .fastq files

If entire flowcell or lane is purchased, the raw data can be delivered as BCL files if requested at time of contract

Why Our Clients Choose Psomagen


Superior Data Quality

Exceeds manufacturer benchmarks


Fast Turnaround

Starting at one week for sequencing projects


US-Based Processing

Sequencing and customer support in Rockville, MD


Ph.D. Level Support

Expert assistance every step of the way, with real-time project updates through our online system


Automated Workflows

Increase scalability and improve reproducibility


CLIA-Certified, CAP-Accredited Labs

Clinical research options available (inquire for more information)


Complete Sequencing Solutions

From extraction to data analysis, with many sample types accepted

Quotes From Our Clients

I chose Psomagen because of the price and US-based labs. We've used Psomagen before and were very satisfied. The people were responsive and helpful and the work done was very good.


University Research Laboratory

I chose Psomagen because of the price and US-based labs. We've used Psomagen before and were very satisfied. The people were responsive and helpful and the work done was very good.


University Research Laboratory

I chose Psomagen because of the price and US-based labs. We've used Psomagen before and were very satisfied. The people were responsive and helpful and the work done was very good.


University Research Laboratory

We're Ready for Your Order!

Here's what to expect in your project's life cycle. If you have a question during sequencing, it's easy to get in touch!


Request a Consultation

Create an account through our client portal and get in touch with your local field team to start designing a project.

We’ll help drive your research forward with the right tools for your research needs.


Place an Order

After finalizing the scope of work, the official quote will be issued for signing. Then, your sales representative will send specific shipping instructions.

Once all the samples and necessary documents are ready, we can start your project. Need more analysis? Reach out to our bioinformatics experts to dig deeper.


QC and Data Delivery

All Psomagen sequencing projects come with a built-in QC step to ensure you get the information you need. An account manager will update you on your project's progress in a timely manner.

You can also see some project information and securely view all valid reports on the client portal site.

Customer Publications and Other Resources

A chromosome-level reference genome and pangenome for barn swallow population genomics
Whole genome sequencing identifies structural variants contributing to hematologic traits in the NHLBI TOPMed program
First complete mitochondrial genome of the Saharan striped polecat (Ictonyx libycus)
PSO Comprehensive!

What forms of RNA sequencing will get you the insights you need?