Exosomal microRNA Sequencing

Exosomal microRNA (miRNA) sequencing is a next generation sequencing (NGS)-based method to comprehensively detect and novel exosomal miRNAs. It can quickly identify the complete picture of the miRNAs in exosomes from different sources. Our experienced technical team provides you with sample-to-data exosomal miRNA sequencing and comprehensive bioinformatics analysis.

Overview

The exosome is a vesicle secreted by eukaryotic cells with lipid bilayer membrane of about 30-150 nm in diameter. Exosomes contain cell-specific proteins, lipids and nucleic acids, which can be transferred to other cells as signal molecules to change the function of other cells and participate in pathological processes such as immune response, antigen presentation, tumor metastasis, and nervous system. miRNA is a type of non-coding RNA of about 22nt in length. These small RNAs usually exist in body fluids such as exosomes and guide the silencing complex (RISC) to degrade mRNA or inhibit translation of mRNA through a complementary base pairing with the target mRNA. The lipid bilayer structure of exosomes can protect miRNA well, and the expression level of exosomal miRNA significantly varies with exosomal sources and physiological conditions.

We provide mature exosomal microRNA sequencing technology including sample QC, exosomal miRNA library construction, high-throughput sequencing based on Illumina HiSeq and detailed bioinformatics analysis. Millions of miRNA sequences can be obtained at one time, enabling quick identification and quantification of known and unknown exosomal miRNAs and discovery of exosomal miRNAs that are differentially expressed in different tissues, developmental stages, or disease states. Meanwhile, this technology serves as a powerful tool to study the role of exosomal miRNAs in cellular processes, and increase our understanding of the biological mechanisms of tumorigenesis, growth, invasion, and metastasis.

Features

High Resolution Deep Sequencing Strict Quality Control Flexibility
Accurately identify base variations in miRNAs and other small RNAs. More than five million sequences can be obtained at one time. Quality control at every stage from library construction to bioinformatics analysis. Provide one-stop, custom exosomal miRNA research solution.

Project Workflow

Sample Preparation

1. Sample Preparation

Quality assessment and quantification;
RNA purification.

Library Preparation

2. Library Preparation

Construct small RNA cDNA libraries.

Sequencing

3. Sequencing

Illumina HiSeq;
PE50/75/100/150;
10M raw data.

Data Analysis

4. Data Analysis

Visualize and preprocess results and perform custom bioinformatics analysis.

Bioinformatics Analysis Pipeline

Exosomal microRNA Sequencing

In-depth data analysis:

  • miRNA sequence length distribution
  • Differential expression analysis of miRNA
  • Predict novel miRNA
  • Target gene prediction of known miRNA and novel miRNA
  • GO annotation analysis
  • KEGG pathway enrichment analysis
  • SNP analysis of known miRNA
  • Association analysis of miRNA and mRNA

Sample Requirements

RNA sample quantity ≥ 2 ug.

OD260/280 ≥ 1.8, OD260/230 ≥ 1.5, RIN ≥ 7.0, 28S:18S ≥ 1.5.

Please make sure that the RNA is not significantly degraded.

Sample storage: RNA can be dissolved in ethanol 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:

  1. Rabinowits G, Gerçel-Taylor C, Day JM, et al. Exosomal microRNA: a diagnostic marker for lung cancer. Clin Lung Cancer, 2009, 10(1): 42-46.
  2. Bhome R, Del Vecchio F, Lee G H, et al. Exosomal microRNAs (exomiRs): Small molecules with a big role in cancer. Cancer Lett, 2018, 4, 28 (420): 228-235.
  3. Sun Z, Shi K, Yang S, et al. Effect of exosomal miRNA on cancer biology and clinical applications. Mol Cancer, 2018, 11,17(1): 147-153.
  4. Sohn W, Kim J, Kang S H, et al. Serum exosomal microRNAs as novel biomarkers for hepatocellular carcinoma. Exp Mol Med, 2015, 18, 47: e184.
* For Research Use Only. Not for use in diagnostic procedures.


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