Small RNA Sequencing
Paving the path for NGS : Biomarker Discovery
Small RNA (microRNA) Sequencing
- Capture a Complete Range of Small RNA and miRNA species
- Sequence Ultra-Low Inputs from Liquid Biopsies and Exosomes
- Illumina® NextSeq Sequencing Platform
- Understand Post-Transcriptional Regulation with Bioinformatics Analysis Service
- Examine Differential Expression and Discover Novel Biomarkers
- Fast Turn Around Time | 3 - 4 weeks
In our accredited state-of-the-art laboratory, we offer comprehensive Next-Generation Sequencing (NGS) services to provide full workflow solutions, from RNA isolation and sequencing to advanced bioinformatic analyses for ready-to-publish data. Our experienced staff have extensive expertise working with a variety of sample types, including, urine, tissue, exosomes, plasma/serum, saliva, FFPE, cerebral spinal fluid (CSF) and more. We are here to help!
Bioinformatics Analysis
Figure 1. (Advanced Analysis) Heat map displaying the hierarchal clustering of miRNAs and samples based on their expression levels. The graph was generated by using 50 miRNAs that have the highest %CV. The color scale indicates the relative expression level of a miRNA to the mean, where green and red indicate higher or lower expression, respectively.
Figure 1. (Advanced Analysis) Heat map displaying the hierarchal clustering of miRNAs and samples based on their expression levels. The graph was generated by using 50 miRNAs that have the highest %CV. The color scale indicates the relative expression level of a miRNA to the mean, where green and red indicate higher or lower expression, respectively.
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Figure 2. Graph illustrating that the relative expression level of each microRNA detected is highly correlated between 50µL and 4mL of plasma.
Figure 2. Graph illustrating that the relative expression level of each microRNA detected is highly correlated between 50µL and 4mL of plasma.
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Figure 3. Graph illustrating that the relative expression level of each microRNA detected is highly correlated between 200µL and 4mL of plasma.
Figure 3. Graph illustrating that the relative expression level of each microRNA detected is highly correlated between 200µL and 4mL of plasma.
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Figure 4. (Advanced Analysis) Bubble plot illustrating GO terms determined to be significantly enriched within the up-regulated differentially expressed genes. GO terms that are significantly enriched are displayed as their GO:ID on the plot, but their associated term is listed and grouped by their respective domains.
Figure 4. (Advanced Analysis) Bubble plot illustrating GO terms determined to be significantly enriched within the up-regulated differentially expressed genes. GO terms that are significantly enriched are displayed as their GO:ID on the plot, but their associated term is listed and grouped by their respective domains.
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Figure 5. (Advanced Analysis) PCA (Principle Component Analysis) illustrating the distance between samples based on their miRNA profiles. The plot was generated by using 50 miRNAs which show the highest variation across all samples, calculated by the %CV (coefficient of variation).
Figure 5. (Advanced Analysis) PCA (Principle Component Analysis) illustrating the distance between samples based on their miRNA profiles. The plot was generated by using 50 miRNAs which show the highest variation across all samples, calculated by the %CV (coefficient of variation).
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Figure 6 Graph illustrating the number of microRNAs detected from 50 µL, 200 µL and 4mL of plasma. The numbers of miRNAs from varying volumes do not differ significantly.
Figure 6 Graph illustrating the number of microRNAs detected from 50 µL, 200 µL and 4mL of plasma. The numbers of miRNAs from varying volumes do not differ significantly.
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Figure 7. Graph showing different small RNA biotype fractions within each sample as well as between sample relatively to the number of mapped reads.
Figure 7. Graph showing different small RNA biotype fractions within each sample as well as between sample relatively to the number of mapped reads.
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Figure 8. (Advanced Analysis) Venn diagram showing the different numbers of miRNA detected in different volumes of plasma. The diagram illustrates that of the microRNAs identified, the majority are detected in all volumes of plasma input from 50 μL to 4 mL.
Figure 8. (Advanced Analysis) Venn diagram showing the different numbers of miRNA detected in different volumes of plasma. The diagram illustrates that of the microRNAs identified, the majority are detected in all volumes of plasma input from 50 μL to 4 mL.
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Figure 9. Volcano plot illustrating the relation between –log10 FDR and log2 fold change between the control and treatment groups. Highly deregulated miRNAs have a fold change of ≥2 (log2 fold change of ≥1 or ≤-1) while significantly deregulated miRNAs have an FDR of ≤0.05 (-log10 FDR of ≥1.30103).
Figure 9. Volcano plot illustrating the relation between –log10 FDR and log2 fold change between the control and treatment groups. Highly deregulated miRNAs have a fold change of ≥2 (log2 fold change of ≥1 or ≤-1) while significantly deregulated miRNAs have an FDR of ≤0.05 (-log10 FDR of ≥1.30103).
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Your Custom Workflow
1. Consultation
2. Sample Submission/Shipping
3. Sample Isolation
4. RNA Quality Control
5. Library Preparation
6. Sequencing
7. Analysis
8. Final Report
Step 1: Consultation
Get started by requesting your free consultation with one of our NGS experts. We would love to discuss your research objectives and are happy to provide assistance in defining your project goals. Our NGS specialists and bioinformaticians are here to provide customized recommendations to suit your project needs.
Request Your Consultation
Step 2: Sample Submission & Shipping
Shipping your samples to our facility is easy and painless! We’ll provide you with detailed instructions on how to handle, process and ship your samples to our laboratory. We accept both purified RNA and specimens for RNA isolation services.
If you are shipping specimens to us, a complete guide on sample collection and preservation will be provided to you. Recommended specimen input and handling procedures for RNA purification can be found in our Small RNA-Seq Service Guide.
How will you submit your samples?
For detailed shipping instructions, please refer to the document below.
Shipping Instructions
Step 3: Sample Isolation
Our team has tremendous experience and expertise in isolating RNA from ultra-low concentration samples such as liquid biopsies (plasma, serum, urine, CSF), exosomes and FFPE. Utilizing our proprietary silicon carbide resin for all isolations allows us to capture miRNA with greater diversity and without the use of phenol.
Step 4: RNA Quality Control
You can be confident that the sequencing portion of your project will be successful as we perform quality control on the isolated RNA to assess quantity, quality and amplifiability prior to library preparation.
Step 5: Library Preparation
Our team has expertise in RNA library preparation from all types of specimens, successfully generating libraries from ultra-low RNA inputs. We will generate the small RNA libraries using Norgen Biotek’s Small RNA Library Preparation Kit (for Illumina). The libraries will then undergo a quality check prior to sequencing.
Step 6: Sequencing
We will sequence the sample libraries on the Illumina® NextSeq platform to achieve your required coverage and read depth.
Step 7: Bioinformatics Analysis
Once the sequencing is complete, our team of bioinformaticians conduct a comprehensive analysis to help make sense of your data. Customized advanced analyses, such as Differential Expression, KEGG/GO and Novel miRNA Discovery can be added to the service to answer a specific biological question. You will receive a final report, which will include a summary of your project as well as your data illustrated in ready-to-publish figures.
Sample Report
Let's get your project started.
Request Your ConsultationQuality In → Quality Out
We believe that the best research starts with quality sample collection and preservation.
Saliva RNA Collection and Preservation Devices Dx
(Cat. 53800)
CE-IVD marked in accordance with EU Directive 98/79/EC
Total Nucleic Acid Preservation Tubes Dx
(Cat. Dx69200)
CE-IVD marked in accordance with EU Directive 98/79/EC
Stool Nucleic Acid Collection and Preservation Devices Dx
(Cat. Dx45660)
CE-IVD marked in accordance with EU Directive 98/79/EC
Stool Nucleic Acid Collection and Preservation System
(Cat. 63700)
For Research Use
Fecal Swab Collection and Preservation System
(Cat. 45670)
For Research Use
Urine Preservation
(Cat. 18116, 18118, 18120)
For Research Use
Urine Collection and Preservation Cup
(Cat. 18129)
For Research Use
cf-DNA/cf-RNA Preservative Tubes Dx
(Cat. Dx63950)
CE-IVD marked in accordance with EU Directive 98/79/EC
Frequently Asked Questions
What is Next Generation Sequencing?
Next Generation Sequencing (NGS) is an important tool used in the field of genomic research as it allows for high throughput and massively parallel sequencing of thousands of molecules simultaneously. NGS provides researchers with fast, scalable solutions for a clear, complete picture of their samples to advance their scientific discoveries.
What is bioinformatics?
Bioinformatics has quickly advanced to become an important and powerful application in biological research. It involves a combination of computational and statistical methods to better understand and analyze scientific data, guiding the research community to new opportunities and novel discoveries. As an emerging discipline, bioinformatics continues to be an essential tool for researchers in many fields including modern molecular biology and clinical research, with its applications leading to significant medical advancements and developments in future research to come.
Do you offer extraction services?
Yes. RNA extraction service can be included for any RNA-Seq project at an additional cost.
What are your sample submission requirements?
Our requirements for sample submission can be found under Step 2: Sample Submission on our small RNA NGS Service page.
What sequencing platform do you sequence on?
We sequence all RNA Sequencing projects on the Illumina® NextSeq platform.
What is the depth of coverage Norgen provides?
Our standard depth is 10 Million (average) raw reads per sample. We can accommodate higher or lower read depth if required. Please contact services@norgenbiotek.com to discuss the optimal read depth for your project goals.
Does the price include bioinformatics analysis?
Yes. Our comprehensive service includes a basic bioinformatics analysis of your raw data. We deliver the bioinformatics analysis in a PDF report and we also provide you with a secure link to download all of your raw data (fastq) and intermediate files. In order to get the advanced analysis for Small RNA Sequencing, there is an additional fee.
What is the typical turnaround time?
Our typical turnaround time is 3-4 weeks from the time of sample receipt. The turnaround time is dependent on the size of your project. Please contact services@norgenbiotek.com to receive an accurate estimated turnaround time.
Norgen is located in Canada. Do you still accept samples from other countries?
Absolutely! We accept samples from countries worldwide and our NGS team will be happy to assist you with making the shipping arrangements.
Citations
| Title | Small RNA-sequence analysis of plasma-derived extracellular vesicle miRNAs in smokers and patients with chronic obstructive pulmonary disease as circulating biomarkers |
| Journal | Journal of Extracellular Vesicles. 2019. |
| Authors | Isaac Kirubakaran Sundar, Dongmei Li, Irfan Rahman |
| Title | Circulating MicroRNAs and Treatment Response in Childhood Asthma |
| Journal | American Journal of Respiratory and Critical Care Medicine. 2020. |
| Authors | Jiang Li, Ronald Panganiban, Alvin T. Kho, Michael J. McGeachie, Leanna Farnam, Robert P. Chase, Scott T. Weiss, Quan Lu, and Kelan G. Tantisira |
| Title | Extracellular vesicles from endometriosis patients are characterized by a unique miRNA-lncRNA signature |
| Journal | JCI Insight. 2019. |
| Authors | Kasra Khalaj, Jessica E Miller, Harshavardhan Lingegowda, Asgerally T Fazleabas, Steven L Young, Bruce A Lessey, Madhuri Koti, Chandrakant Tayade |
| Title | Exosomal microRNAs are novel circulating biomarkers in cigarette, waterpipe smokers, E-cigarette users and dual smokers |
| Journal | BMC Medical Genomics. 2020. |
| Authors | Kameshwar P. Singh, Krishna P. Maremanda, Dongmei Li & Irfan Rahman |
| Title | Urine miRNAs as potential biomarkers for systemic reactions induced by exposure to embedded metal |
| Journal | Biomarkers in Medicine. 2021. |
| Authors | Ivan J Vechetti , Yuan Wen, Jessica F Hoffman, Alexander P Alimov, Vernieda B Vergara, John F Kalinich, Joanna M Gaitens, Stella E Hines, Melissa A McDiarmid, John J McCarthy & Charlotte A Peterson |
| Title | High Throughput Sequencing of MicroRNA in Rainbow Trout Plasma, Mucus, and Surrounding Water Following Acute Stress |
| Journal | Frontiers in Physiology. 2021. |
| Authors | Heather Ikert, Michael D. J. Lynch, Andrew C. Doxey, John P. Giesy, Mark R. Servos, Barbara A. Katzenback, Paul M. Craige |
| Title | Distinct Exosomal miRNA Profiles from BALF and Lung Tissue of COPD and IPF Patients |
| Journal | International Journal of Molecular Sciences. 2021. |
| Authors | Gagandeep Kaur, Krishna Prahlad Maremanda, Michael Campos, Hitendra S. Chand, Feng Li, Nikhil Hirani, M. A. Haseeb, Dongmei Li, Irfan Rahman |
Our NGS Clients
We are pleased to work with some of the most prestigious companies and institutions in 155+ countries.
