Exosomal Small RNA Sequencing

Exosomal small RNA sequencing is a next-generation sequencing (NGS)-based method for comprehensive transcriptome analysis of exosomal small RNA species. It can accurately explore the mechanisms underlying gene expression regulation and intercellular communication, and discover biomarkers for disease diagnosis and monitoring. We provide high-throughput sequencing of exosomal small RNAs to quickly and efficiently obtain the global information of exosomal small RNAs.

Overview

The exosome is a type of membranous vesicles produced in the endosomal compartment of eukaryotic cells and releases its vesicular cargo into the extracellular space. It is typically 30-100 nm in diameter and contains nucleic acids and proteins. Exosomes serve as transporters of these substances between cells and participate in physiological processes such as immune response, inflammatory response, angiogenesis, apoptosis, coagulation, and waste disposal. It can be used as an early diagnostic marker for diseases and as a carrier for drug delivery and targeted drug therapy. Small RNAs are non-coding RNA (ncRNA) molecules with length less than 200 nucleotides, mainly including microRNA, tRF, tiRNA, piRNA, tRNA, snoRNA and pre-microRNA. As important regulatory molecules, small RNAs are involved in gene expression, epigenetics, and development of diseases. The expression of exosomal small RNA significantly varies in different disease models.

Exosomal small RNA sequencing (RNA-seq) can quickly and efficiently obtain comprehensive information of exosomal small RNAs, enabling target gene analysis and functional analysis. Exosomal small RNA-seq is an ideal method for the discovery of biomarkers as its convenience for identifying the differential expression patterns in disease models. We provide customers with comprehensive exosomal small RNA-seq service and in-depth data analysis for a better understanding of the roles that exosome small RNAs play in gene expression regulation and intercellular communication, so as to identify potential therapeutic targets, early diagnostic biomarkers and targeted drug carriers for multiple diseases.

Features

High Resolution Comprehensive Information Accurate Annotation Superiority
Allow single-base resolution identification of small RNAs. Detect multiple types of small RNAs at one time, including miRNA, siRNA, piRNA, etc. Provide complete annotate information with multiple databases for exosomes or vesicles. Rich experience in experimental design, operations and data analysis.

Project Workflow

Exosomal Small RNA Sequencing

1. Sample Preparation

Quality assessment and quantification.

Exosomal Small RNA Sequencing

2. Library Preparation

Size selection (17 to 35 nt), cDNA library construction, library QC by BioAnalyzer and RTQ-PCR.

Exosomal Small RNA Sequencing

3. Sequencing

Illumina HiSeq;
50/75/100 nt, single or paired-end.

Exosomal Small RNA Sequencing

4. Data Analysis

Perform custom bioinformatics analyses and provide detailed data report.

Bioinformatics Analysis Pipeline

Exosomal Small RNA Sequencing

In-depth data analysis:

  • Length distribution of small RNA
  • Quantitative analysis of small RNA
  • Differential expression analysis of small RNA
  • Predict novel small RNA
  • Target gene prediction of miRNA
  • GO annotation analysis
  • KEGG pathway enrichment analysis
  • Cluster analysis
  • Interactive network analysis

Sample Requirements

RNA sample quantity ≥ 2 ng, volume ≥ 20ul.

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. 1. Zhu L, Li J, Gong Y, et al. Exosomal tRNA-derived small RNA as a promising biomarker for cancer diagnosis. Mol Cancer, 2019, 2, 18(1): 74.
  2. 2. Shurtleff M J, Yao J, Qin Y, et al. Broad role for YBX1 in defining the small noncoding RNA composition of exosomes. Proc Natl Acad Sci U S A, 2017, 24, 114(43): E8987-E8995.
  3. 3. Lunavat T R, Cheng L, Kim D K, et al. Small RNA deep sequencing discriminates subsets of extracellular vesicles released by melanoma cells--Evidence of unique microRNA cargos. RNA Biol, 2015, 12(8): 810-823.
* For Research Use Only. Not for use in diagnostic procedures.


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