Small Extracellular Vesicles/Exosomes Transcriptomics

Small Extracellular Vesicles/Exosomes Transcriptomics

Exosome RNA, comprising various types such as mRNA, miRNA, and IncRNA ,ects.,plays a significant role in biological functions. Exosome RNA sequencing is to obtain RNA sequence information by sequencing exosome RNA, so as to analyze and study RNA. This technology has been widely applied in research on diverse diseases, including cancer, neurodegenerative diseases, and cardiovascular diseases. Exosomal RNA serves as a crucial biomarker, providing important scientific basis for the diagnosis, treatment and prognosis of diseases. Services related to exosomal RNA sequencing typically encompass multiple stages, include exosome RNA extraction, sequencing, and data analysis. Through these stages, researchers can obtain comprehensive information about exosomal RNA, including RNA expression levels, splicing variations, post-transcriptional modifications and other information, and then analyze and study the function of RNA.

Small RNA Sequencing

Small RNA mainly includes RNA molecules such as miRNA, piRNA, tsRNA, and SnoRNA. Small RNA is an indispensable regulatory factor in life activities, which participates in important biological processes such as gene expression and regulation, RNA processing and splicing, protein translation, genetic “invasion” inhibition, and gametogenesis, etc. MicroRNA (miRNA) is the most important part of small RNA and has been proven to be closely related to a variety of diseases including cancer, cardiovascular diseases, and immune system diseases. In addition, exosomal miRNA can also serve as biomarker for disease and is widely used in fields such as disease diagnosis, personalized treatment and prognosis. High-throughput sequencing technologies enable the acquisition of sequence information for all small RNAs in a single sequencing effort. This allows for the analysis of small RNA expression levels under different disease states, sample sources, and treatment conditions, and the precise comparison of small RNA expression differences between them. This, in turn, provides guidance for subsequent studies on the biological significance of small RNAs.

NextGen Exsome employs a library construction system based on QIAseq, which can complete the construction of a miRNA library with as little as 1ng of total RNA, and the optimized chemistry reaction can achieve the construction of a stable miRNA-specific library, minimize reaction deviations and eliminate adapter dimers. Unique molecular identifier (UMI) mark each miRNA at an early stage, eliminating PCR and sequencing bias. This makes it a relatively high-quality library construction approach for miRNA from exosomal samples in terms of data quality.

Non-coding RNA Sequencing

Exosomal RNA, especially non-coding RNA, has obvious expression differences in different disease models and is expected to become an important molecular marker for liquid biopsy and reveal the mechanism of disease. High-throughput transcriptome sequencing technology enables the simultaneous detection of various RNA molecules such as lncRNA, mRNA, and circRNA, facilitating research into the identification and regulatory functions of exosomal non-coding RNA molecules.

NextGen Exsome utilizes a SMARTer-based library construction system, requiring as little as 1ng of total RNA to complete lncRNA library preparation. Subsequent sequencing data can be analyzed separately for LncRNA and mRNA, or combined with miRNA sequencing data for full transcriptome analysis (including LncRNA, mRNA, CircRNA, and miRNA).

Services at NextGen Exsome

1.Exosomes RNA Extraction

Using exosome-specific micro-RNA extraction kit to extract RNA and detect the concentration.

Deliverables:

RNA concentration data in XLSX format

2.Exosomes RNA Quality Control

Using capillary electrophoresis to detect the fragment distribution of nucleic acids.

Deliverables:

1)Sample analysis report in PDF format

2)Instrument raw data in BOPX format

3.Exosomes miRNA Sequencing

The workflow includes library construction quality control, on-machine sequencing, and standard analysis. It utilizes a micro-library preparation approach, with Illumina platform PE150 sequencing strategy. The data output is 10M reads (3G Rawdata).