Exosomes are small (30-150 nm) vesicles containing unique RNA and protein cargo, 
secreted by all cell types in culture. They are also found in abundance in body 
fluids including blood, saliva, and urine. At the moment, the mechanism of 
exosome formation, the makeup of the cargo, biological pathways, and resulting 
functions are incompletely understood. One of their most intriguing roles is 
intercellular communication--exosomes function as the messengers, delivering 
various effector or signaling macromolecules between specific cells. There is an 
exponentially growing need to dissect structure and the function of exosomes and 
utilize them for development of minimally invasive diagnostics and therapeutics. 
Critical to further our understanding of exosomes is the development of 
reagents, tools, and protocols for their isolation, characterization, and 
analysis of their RNA and protein contents. Here we describe a complete exosome 
workflow solution, starting from fast and efficient extraction of exosomes from 
cell culture media and serum to isolation of RNA followed by characterization of 
exosomal RNA content using qRT-PCR and next-generation sequencing techniques. 
Effectiveness of this workflow is exemplified by analysis of the RNA content of 
exosomes derived from HeLa cell culture media and human serum, using Ion Torrent 
PGM as a sequencing platform.