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Omics Research // Transcriptomics

Low-Input RNA Sequencing

Use a small amount of RNA input to accurately measure the number of genes and transcripts in all coding and non-coding RNA molecules within the transcriptome.

Let's Talk About Low-Input RNA Seq

What Is Low-Input RNA Sequencing?

Low-input RNAseq assesses the transcriptome in challenging sample types where there is a small amount of cells, tissue, or RNA available.

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Why Is Low-Input RNA Sequencing Important?

Low-input RNAseq enables researchers to analyze gene expression and transcription activities within cells they otherwise would not be able to use due to limitation of available RNA.

This becomes important in assessing the mechanisms of rare subtypes of cell populations in oncology, neuroscience, and targets for treatment.

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What Can You Do?
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Template Switching

RNA sequencing using less than 10 ng by eliminating litigation using a template switching mechanism

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Strand-Specific Sequencing

Identify which strand of DNA the transcript was derived from

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High-Sensitivity Results

Sensitive as a result of efficient removal of cDNA derived from ribosomal RNA (rRNA) rather than an rRNA depletion process

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Experienced Bioinformatics Experts

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

Low-Input RNA Sequencing Technologies

Switching Mechanism Technology

Improve your sequencing performance using technology that uses a switching mechanism at the 5’ end of the RNA template.

Depletion of cDNA Derived from rRNA

Removal of cDNA derived from rRNA eliminates the majority of the high abundance rRNA that yields high mappable sequencing coverage. This negative selection type approach increases the amount of relevant sequencing reads and examines:

  • 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.

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Total RNA Sequencing Best Practices

Sample Preparation & RNA Extraction

RNA extraction from cells, tissue, and FFPE samples*

  • Sample QC: >10 ng in nuclease-free water

*Low-input RNA sequencing is an alternative approach for tissue stored as formalin-fixed, paraffin-embedded (FFPE) blocks, as some of the RNA may be degraded and in small amounts.

Library Preparation

  • SMARTer Stranded RNA library
  • SMARTer® Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian

Sequencing Instrumentation & Read Length

Illumina NovaSeq 6000

  • 2 x 151 bp read length

If you are interested in assessing novel alternative transcripts and RNA editing, we recommend a target read sequencing 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:

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

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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

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More Reviews on SelectScience®

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

5-star
More Reviews on SelectScience®

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

5-star
More Reviews on SelectScience®

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!

01

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

More Customer Publications
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 Much More RNA Sequencing

mRNA Sequencing

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

Single-Cell RNA Sequencing

Analyze transcriptional changes on a cell-by-cell basis.

Sequence cellular diversity within a heterogeneous matrix for gene expression and precision medicine.

Total RNA Sequencing

For the most comprehensive data on the whole transcriptome. Encompass all RNA types and sizes recoverable via total RNA sequencing.

PSO Comprehensive!

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

Ask Our Experts