Exosomal Long RNA-seq

CD Genomics’s exosomal long RNA sequencing (RNA-seq) service enables comprehensive analysis of mRNA, circRNAs, and lncRNAs in exosomes, offering the opportunity to quantify expression levels of individual transcript isoforms, analyze the mechanisms underlying human diseases, and discover potential biomarkers. Our project manager will be in close communication with customers, from early consultations to and post-delivery assistance.

Overview

Exosomes secreted by most cell types are lipid-rich small vesicles that contain and transport biomolecules such as RNAs and proteins between cells and sometimes serve as a platform for cell-cell communication and disease transmission. Therefore, exosomes are now progressively considered to be potential sources for circulating early diagnostic biomarkers. So far, most studies have concentrated on the changes of microRNA in circulating exosomes, however, more and more evidence indicates that long RNAs consisting of circular RNAs (circRNAs), long noncoding RNAs (lncRNAs), and messenger RNA (mRNA) may have functional and potential clinical implications.

Exosomal Long RNA-seq is the method that combines exosomes isolation and RNA sequencing to profile the circulating exosomal long RNAs (exoLRs) and explore the alteration of exoLRs. It might contribute to promoting the investigation of the mechanisms of human diseases and discovery of potential biomarkers for clinical diagnosis. CD Genomics provides quality assessment of the purified exosomal RNAs by real-time PCR and electrophoresis. Subsequently, ribosomal RNA are depleted and stranded-specific DNA libraries are generated, followed by high-throughput sequencing and data analysis. We provide differential expression, statistical analyses, and custom analyses to support exosomal long RNA characterization and the development of companion diagnostics and other applications.

Features

Strand-specific libraries Transcriptome-Wide Bioinformatics Analysis One-Stop Solution
Ribosomal-depleted strand-specific RNA-seq libraries containing both polyA+ and polyA- sequences. Profile all mRNAs, lncRNAs and circRNAs in exosomal samples. Data preprocessing, RNA prediction, differential expression analysis, etc. One-stop solution from sample QC and library construction, to sequencing and data analysis.

Project Workflow

Sample Preparation

1. Sample Preparation

Stringent sample QC,
rRNA removal.

Library Preparation

2. Library Preparation

Strand-specific libraries are constructed, and assessed and quantified by BioAnalyzer and RTQ-PCR.

Sequencing

3. Sequencing

HiSeq X10/4000;
PE50/75/100/150;
>10G clean data

Data Analysis

4. Data Analysis

Visualize and preprocess results, and perform custom bioinformatics analysis.

Bioinformatics Analysis Pipeline

Exosomal Long RNA-seq

In-depth data analysis:

  • Genome mapping
  • Profiling of lncRNA, circRNA and mRNA
  • Prediction of novel lncRNA, circRNA and mRNA
  • Detection of novel and rare transcript variants
  • Differential expression analysis
  • Target gene prediction and functional analysis
  • Gene ontology and KEGG enrichment analysis
  • Annotation of circRNA and lncRNA

Sample Requirements

Total RNA (concentration ≥ 1 ng/uL, quantity ≥ 20 ng) from human tumor and adjacent normal tissues

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. HE G, HUANG Y, Lin LIU, et al. The alteration of circulating exosomal mRNAs in acute myocardial infarction is associated with inflammation response mediated by neutrophils. 2020.
* For Research Use Only. Not for use in diagnostic procedures.


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