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Omics Research // Transcriptomics

mRNA Sequencing

Use the poly-A tail of mRNA transcripts, positively selecting mRNA out of all the background RNA, to isolate the coding RNA sequences of actively transcribed genes. Obtain functional analysis of transcriptional regulators in disease and baseline biological processes.

What Is mRNA Sequencing?

mRNA sequencing is the method by which coding RNA in the form of messenger RNA (mRNA) is selected by the poly-A tail and sequenced using next-generation sequencing.

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Why Is mRNA Sequencing Important?

RNA transcription is the link between DNA and protein expression. Using mRNA-seq enables the user to analyze gene expression patterns by sequencing less raw throughput while maintaining high mappable sequencing coverage.

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.

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What Can You Do?
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Positive Selection

mRNA selection using the poly-A tail of extracted RNA

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Strand-Specific Sequencing

Identify which strand of DNA the transcript was derived from

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High Specificity

Positive selection leads to uniform coverage of RNA that codes for proteins and other polyadenylated non-coding RNA

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Experienced Bioinformatics Experts

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

Technologies Used for mRNA Sequencing

Stranded mRNA Seq

Psomagen uses strand-specific mRNA sequencing, or stranded mRNAseq, to identify overlapping transcripts and genes.

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

Gene Expression Profiling

Gene expression profiling analyzes known genes. Transcripts are assembled with StringTie software based on a reference genome model.

After assembly, the abundance of gene/transcript is calculated in the read count and normalized value as FPKM (Fragments Per Kilobase of transcript per Million mapped reads) for each sample. Isoforms of known transcripts are provided.

On-Going Promotions

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

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mRNA 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 mRNA
  • 500 ng Total RNA (25-1000 ng)
  • Library QC >10 nM at ~260 bp

Sequencing Instrumentation & Read Length

Illumina NovaSeq 6000

  • 2 x 150 bp read length
  • Custom read 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 the entire flowcell or lane is purchased, the raw data can be delivered as BCL files if requested at the time of contract.

Why Our Clients Choose Psomagen

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Superior Data Quality

Exceeds manufacturer benchmarks

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Fast Turnaround

Starting at one week for sequencing projects

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US-Based Processing

Sequencing and customer support in Rockville, MD

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Ph.D. Level Support

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

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Automated Workflows

Increase scalability and improve reproducibility

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CLIA-Certified, CAP-Accredited Labs

Clinical research options available (inquire for more information)

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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.

Professor

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.

Professor

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.

Professor

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!

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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.

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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.

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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?