Protocols

Protocols

   

The consortium has developed Standard Operating Procedures (SOPs) for purification of extracellular vesicles and RNA, as well as for exRNA data analysis. A major goal of the consortium is to standardize these procedures to minimize experimental variation across labs and experiments.

Many of the protocols have been updated since first release in 2015 based on our experience vetting the originals in multiple labs. The major change is an increase in input volume of biofluid from 200 µL to 500 µL. We found that the larger input volume led to more reproducible amounts of RNA isolation across multiple experiments.

Updated protocols are indicated here as version 2 (v2). We have maintained links to version 1 protocols for archival purposes.

   

Protocols Decision Tree

This decision tree allows a beginner to understand the best approaches available in the field to study extracellular vesicles and extracellular RNA. See this blog post for an in-depth discussion of how to select protocols from this page. The Protocols Decision Tree currently (Nov 2017) still refers to version 1 protocols.

 

Collection of Biofluids

 

Isolation and Enrichment of Extracellular Vesicles

 

RNA Isolation

 

Validation and Quantification

 

RNA Sequencing

 

Proteomics on Extracellular Vesicles