miRNA sequencing, based on next-generation sequencing (NGS), can comprehensively profile miRNA sequences, either known or novel miRNAs. Our miRNA sequencing detects novel miRNAs as well as isomiR, enabling you to see precisely which miRNA sequences are expressed in your samples and uncover the importance of these small regulatory elements linked to a wide range of biological functions.
Overview
The miRNAs, 19 to 25 nucleotides long, are a class of small non-coding RNAs, processed from hairpin precursors. miRNA play a key role for the translation efficiency and the stability of their target mRNAs. The miRNAs can control the expression of up to several hundreds of genes by guiding the RNA-induced silencing complex (RISC) to complementary sites in the 3′UTRs of the target mRNAs. This regulationary mechanism underlying gene expression is involved in almost all processes of cell biology and development, such as cell differentiation and apoptosis, the clearance of maternally deposited transcripts, morphogenesis, and organogenesis.
miRNA sequencing is a technique that examines and quantity and sequences of miRNA molecules in a given sample using NGS and size selection. miRNA sequencing experiments contribute to our understanding of cell biology essential developmental processes by detect both known and novel miRNAs, and how RNA molecules affect gene regulation, thus influencing disease development and phenotypic variation and accelerating the discovery of diagnostic or prognostic biomarkers, as well as the development of miRNA-targeting drugs.
Features
| High Sensitivity | High Precision | Transcriptome Wide | One-Stop Solution |
|---|
| Theoretically detect miRNA at levels down to one copy in a single cell. | Can detect single base change in miRNA. | Profile all miRNAs, either known or unknown, in your biological sample. | From sample QC, library construction, to sequencing and data analysis. |
Project Workflow
1. Sample Preparation
Quality assessment and quantification by locked nucleic acid (LNA) qPCR.
2. Library Preparation
RNA Selection – Size Fractionation; 18~40 bp insert cDNA library; Optimized for biofluids with limited RNA content.
3. Sequencing
50 SE / 75 SE; 7-10 Million Reads.
4. Data Analysis
Data quality is measured, visualize and preprocess results, and perform custom bioinformatics analysis.
Bioinformatics Analysis Pipeline
In-depth data analysis:
- Biostatistical analysis –transcript copy number comparisons, expression analysis, length distribution, multi-parameter data analysis, etc.
- Pre-miRNA clusters
- GO and KEGG enrichment analysis
- Detect novel and rare miRNAs and isomiR
- Identification of differentially expressed miRNAs
- Target gene prediction and functional analysis of miRNAs
- Novel miRNA Prediction
- miRNA structure analysis
Sample Requirements
- RNA sample (concentration ≥ 200 ng/uL, quantity ≥ 2 ug, volume ≥ 5 uL)
- 1.8 ≤ OD260/280 ≤ 2.2, RIN ≥ 6.0, 28S:18S ≥ 1.0.
- Please make sure that the RNA is not degraded.
Sample storage: RNA can be dissolved in ethonal or RNA-free ultra-pure water and stored at -80°C. RNA should avoid repeated freezing and thawing.
Shipping Method: When shipping RNA samples, the RNA sample is stored in a 1.5 mL Eppendorf tube, sealed with sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.
Deliverable: FastQ, BAM, coverage summary, QC report, custom bioinformatics analysis.
References:
- Ninova M, Ronshaugen M, Griffiths-Jones S. MicroRNA evolution, expression, and function during short germband development in Tribolium castaneum. Genome Research, 2016, 13(1): 331.
- Saiselet M, Gacquer D, Spinette A, et al. New global analysis of the microRNA transcriptome of primary tumors and lymph node metastases of papillary thyroid cancer. BMC Genomics, 2015, 16:828.
- Paneru B D, Al-Tobasei R, Kenney B, et al. RNA-Seq reveals MicroRNA expression signature and genetic polymorphism associated with growth and muscle quality traits in rainbow trout. Scientific reports, 2017, 7: 9078.
* For Research Use Only. Not for use in diagnostic procedures.
