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Updated: May 20, 2026
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Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations
Published on: August 4, 2016
Protocol for total RNA sequencing analysis of extracellular RNA from biofluids
Anneleen Decock1, Kimberly Verniers1, Philippe Decruyenaere2
1Department of Biomolecular Medicine, Ghent University, 9000 Ghent, Belgium; OncoRNALab, Cancer Research Institute Ghent (CRIG), 9000 Ghent, Belgium.
STAR Protocols
|May 18, 2026
Summary
This study details a protocol for analyzing extracellular RNAs (exRNAs) in blood plasma using SMART cDNA synthesis technology. This method enables comprehensive RNA sequencing for biomarker discovery in various biofluids.
Area of Science:
- Biochemistry
- Molecular Biology
- Genomics
Background:
- Extracellular RNAs (exRNAs) from biofluids are crucial biomarkers for studying health and disease.
- Current methods for exRNA analysis require standardized protocols for reliable results.
Purpose of the Study:
- To present a detailed protocol for total RNA sequencing of extracellular RNAs (exRNAs) from blood plasma.
- To adapt the protocol for exRNA analysis in other human and animal biofluids.
Main Methods:
- Utilized Switching Mechanism At 5' end of RNA Template (SMART) cDNA synthesis technology for RNA sequencing.
- Described comprehensive steps from blood plasma preparation to sequencing data analysis.
Main Results:
- Successfully established a protocol for blood plasma exRNA sequencing.
- Demonstrated the protocol's applicability to exRNA from various biofluids, including human, murine, and rat samples.
Conclusions:
- The presented protocol provides a robust method for analyzing blood plasma exRNA.
- This protocol facilitates exRNA biomarker discovery across multiple species and biofluid types.
