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.
Why sequence small RNAs?
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.
1. Initial meeting
2. Sample delivery
- 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
4. Library prep/QC and sequencing
- NovaSeq 6000 PE150
- Data required for analyzing smallRNAs: generally 10-20 million reads
5. Data QC
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.
- 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.
We have summarized key information about our Small RNA-Seq service into a service specification sheet.
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