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Next-generation Sequencing

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....

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Updated: Apr 30, 2026

Paper-based Devices for Isolation and Characterization of Extracellular Vesicles

Paper-based Devices for Isolation and Characterization of Extracellular Vesicles

Published on: April 3, 2015

Next Generation Aqueous Two-Phase System for Gentle, Effective, and Timely Extracellular Vesicle Isolation and

Boyang Su^1,2, Morteza Jeyhani^3,4,5, Gobi Thillainadesan¹

¹Biological Sciences Platform, Sunnybrook Research Institute, Toronto, Ontario, Canada.

Journal of Extracellular Vesicles

|March 20, 2025

Summary

This summary is machine-generated.

This study introduces a novel next-generation aqueous two-phase system (next-gen ATPS) for high-purity, high-yield extracellular vesicle (EV) isolation. This method surpasses traditional techniques, improving biomarker analysis and nucleic acid extraction for comprehensive EV research.

Keywords:

EV isolation RNA extraction biomarker analysis circRNA dextranase extracellular vesicles (EVs)miRNA nanoscale flow cytometry next‐generation aqueous two‐phase system (next‐gen ATPS)snoRNA

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Last Updated: Apr 30, 2026

Paper-based Devices for Isolation and Characterization of Extracellular Vesicles

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Published on: April 3, 2015

Purification of High Yield Extracellular Vesicle Preparations Away from Virus

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Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media

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Published on: May 13, 2022

Area of Science:

Biotechnology
Molecular Biology
Cell Biology

Background:

Current extracellular vesicle (EV) isolation methods often sacrifice purity and yield for speed.
Existing techniques like ultracentrifugation (UC) and commercial kits have limitations in efficiency and scalability.

Purpose of the Study:

To develop and validate a next-generation aqueous two-phase system (next-gen ATPS) for superior EV isolation.
To demonstrate the efficacy of next-gen ATPS in enhancing EV purity, yield, and downstream analytical applications.

Main Methods:

Utilized a next-generation aqueous two-phase system (next-gen ATPS) employing polyethylene glycol (PEG) and dextran (DEX) phases.
Isolated EVs by exploiting their preferential migration to the DEX-rich phase.
Incorporated dextranase to facilitate TRIzol RNA isolation and improve immunoreactivity for nanoscale flow cytometry (nFC).

Main Results:

Next-gen ATPS demonstrated superior EV isolation yield and purity compared to UC and commercial kits.
Isolated EVs showed enhanced suitability for biomarker analysis via nFC using pre-conjugated antibodies (CD9, CD63, CD81).
Transcriptomic analysis revealed high overlap in RNA profiles (miRNA, circRNA, snoRNA) with donor cells and other isolation methods, with superior circRNA detection.

Conclusions:

Next-gen ATPS offers a rapid, scalable, and highly effective method for isolating high-quality EVs.
This approach optimizes the extraction and analysis of EV-encapsulated nucleic acids, negating the need for specialized kits.
The method significantly advances EV research by improving isolation efficiency and downstream analysis capabilities.