Exosomal Long RNA Sequencing

Exosomes are tiny membrane-bound extracellular vesicles that are produced in the endosomal compartment of most eukaryotic cells and can be found in many, if not all, body fluids, including blood, urine, and cerebrospinal fluid. And they have emerged as a novel approach for the treatment and diagnosis of various diseases including cancer. Exosomal long RNAs are RNA molecules larger than 200 nucleotides, which mainly include exosomal mRNA, exosomal lncRNA, and exosomal circRNA, which are important mediators of intercellular communication and have become a strong focus of the investigation.

Overview

With the advent of next-generation sequencing technology, long exosome research is accelerating not only in cancer but also in various diseases. It has been found that exosomes. Knowledge of exosome RNA sequencing not only helps to understand their biological roles, but analysis of exosome RNA-Seq data based on bioinformatics can help to find promising biomarkers and diagnostic molecules in the body fluids of patients. Meanwhile, the capability to transfer functionally active cargo highlights their importance as prospective drug delivery systems. Other than miRNA, which has drawn a lot of interests in recent years, exosomal long RNAs, which is comprised of mRNA, lncRNA, and circRNA, may play an equal or perhaps an even more significant role in cell-cell communication by altering biological signaling pathways that affect disease progression, and increasing evidence have shown that the long RNA species have extensive clinical applications.

With decades of experience in high-throughput sequencing, CD Genomics provides high-throughput sequencing services to quickly and accurately obtain the information of exosomal long RNAs. Our one-step exosomal long RNA sequencing services include mRNAs, non-coding RNAs, and circRNAs, enabling alternative splicing analysis and detection of novel transcripts, and gene fusion events. We can generate high-quality, reproducible, and reliable exosomal long RNA libraries and deliver accurate data and comprehensive data analysis using the advanced Illumina next-generation sequencing (NGS) and well-accepted computational tools. Our bioinformatics team provides in-detail long RNA sequencing information interpretation, enabling prediction of long RNA-miRNA-mRNA association network and discovery of molecular markers.

Features

Superb Expertise Transcriptome-Wide Accurate Annotation One-Stop Solution
Decade of experience in high-throughput sequencing and bioinformatics analysis. Profile various exosomal long RNA, including small RNA, microRNA, and lncRNA. Provide complete annotate information with multiple databases for exosomes or vesicles. One-stop solution from sample QC and library construction, to sequencing and data analysis.

Project Workflow

Sample Preparation

1. Sample Preparation

RNA purification;
quality assessment and quantification

Library Preparation

2. Library Preparation

RNA fragmentation;
m7G/m3C library preparation.

Sequencing

3. Sequencing

Illumina HiSeq;
PE 50/75/100/150.

Data Analysis

4. Data Analysis

Visualize and preprocess results, and perform custom bioinformatics analysis.

Bioinformatics Analysis Pipeline

Bioinformatics Analysis Pipeline

In-depth data analysis:

  • Length distribution and quantitative analysis of exosomal long RNA
  • Differential expression analysis of exosomal long RNA
  • Prediction of exosomal long RNA and binding sites
  • Coding-non-coding gene co-expression network (CNC network)
  • GO annotation analysis
  • KEGG pathway analysis
  • Interactive network analysis

Sample Requirements

RNA sample (concentration ≥ 1 ng/uL, quantity ≥ 2 ug)

1.8 ≤ OD260/280 ≤ 2.2, OD260/230≥2.0, RIN ≥ 6.5, 28S:18S ≥ 1.0.

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.

Reference:

  1. Zhou R, Chen KK, Zhang J, et al. The decade of exosomal long RNA species: an emerging cancer antagonist. Molecular cancer. 2018, 17(1):75.
* For Research Use Only. Not for use in diagnostic procedures.


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