Small RNAs regulate gene expression, mainly via RNA silencing. Endogenous expression of microRNA’s (miRNA) can induce the degradation of their complementary messenger RNA, thereby altering overall gene expression levels.
Small RNA-Seq
Detect miRNA and small RNAs in any organism
Why sequence small RNAs?
Discuss your project with me
Small RNAs covered
Small RNA’s are <100 nucleotides in length, and are usually non-coding RNA molecules. GenomeScan’s Small RNA-Seq service allows you to identify and quantify all small RNA types, including miRNA, siRNA, snoRNA, snRNA, mtRNA, piRNA and other non-coding sequences. Our R&D team has developed an accredited method, allowing you to select the size range of the transcripts you are interested in, with great accuracy and robustness.
Low input, robust output
You can start with very little material: the total RNA input can be as low as 10 ng. This opens many possibilities to analyze miRNA in liquid biopsies or tumor biopsies.
Service workflow
1. Initial meeting
When you consider using one of our services, our scientific team will help choosing the optimal project design. Each service can be customized by adding options. You will receive an overview of project budget and deliverables, so you can make an informed choice. Once you return your Purchase Order form, your project manager will contact you to discuss the next steps.
2. Sample delivery
Sample specifications for Small RNA-Seq are:
- Optimal total RNA input: 250ng / sample
- Optimal purified small RNA input: >2 ng / sample
- Ideally deliver ≥250 ng RNA to allow for QC and normalization
- Minimal RNA input: 50 ng. Risk of more adapter content in sequencing data
3. Sample entry QC
After sample arrival, an entry QC is performed to determine if the sample quantity and quality is sufficient so we can guarantee the data outcome stated in your quotation. You will receive a report with sample QC results to inform you of our findings before proceeding further. Samples that fail QC can be replaced by spare samples delivered in the same shipment or sent at a later stage.
4. Library prep/QC and sequencing
The library resulting from sample preparation goes into a library QC step to ensure sufficient material per sample and the right library size. Sequencing on
- NovaSeq 6000 PE150
- Data required for analyzing smallRNAs: generally 10-20 million reads
5. Data QC
The raw data that is generated by the sequencer must be divided to obtain one data file set per sample. This is called demultiplexing.
Your project manager checks the data quality by analyzing the quality metrics of the run. Final inspection of the data takes place and the dataset will be made available via electronic transfer through a secure customer portal or is transferred onto a hard disk.
6. Reporting
You receive your Project Report together with your data. It contains an overview of the experimental methods used, sample specifications, and a summary of all technical information for publication. Deliverables:
- TAT: 3-5 weeks
- Sequencing of small RNAs (raw data)
- Quality score Q30 of ≥80%
- Counts of miRNA (optional)
Cancer research and miRNAs
In cancer research, miRNA’s are often the target of investigation. miRNA-deregulation can lead to all sorts of cancer types, where they are referred to as ‘oncomiRs’.
OncomiRs can also play a role in diagnosis of certain subtypes of cancers, prediction of the disease prognosis, and in stratifying patients. Even in the treatment of certain cancers, mRNA’s can be used as either a tool or a target for therapy.
In certain samples, several miRNA’s might be abundant, thereby masking the detection of other miRNA’s. Our scientists can design custom blocking oligo’s to remove these uninformative miRNAs from the library, so that less sequencing-space is needed.
Service specifications
We have summarized key information about our Small RNA-Seq service into a service specification sheet.
Let's get the conversation started for your next NGS project
Please either fill in this form or email us directly at info@genomescan.nl
and we will get in touch with you to discuss your requirements.
