
Analysis of RNA yield in extracellular vesicles isolated by membrane affinity column and differential ultracentrifugation
Author(s) -
Gilberto Gutiérrez García,
Gabriela Garcı́a,
Jessica Zalapa Soto,
Andrea Medina,
Mariana Rotzinger-Rodríguez,
Gustavo Antonio Casas Aguilar,
Cynthia Paola López Pacheco,
Álvaro Aguayo,
María Montserrat Aguilar-Hernández
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0238545
Subject(s) - rnase p , rna , nucleic acid , ultracentrifuge , yield (engineering) , vesicle , chemistry , rna extraction , chromatography , proteinase k , biochemistry , membrane , biology , microbiology and biotechnology , enzyme , gene , materials science , metallurgy
Extracellular vesicles (EV) have attracted much attention as potential biomarkers due to their protein, RNA and other nucleic acid content. The most common method used for EV isolation is differential ultracentrifugation (DU), however given the DU technical difficulties, other more practical methods have surged, such as membrane-affinity column commercial kits. Here, we assessed one commercial kit in terms of EV recovery and EV-derived RNA yield and compared it with a DU protocol. Our data shows that the commercial kit preparation results in a lower count of EV-like structures and a reduced expression of EV markers when compared to DU samples. Thus, apparently suggesting that the commercial kit had a lower EV yield. However, these findings did not reflect on RNA yield, which was greater with the commercial kit, even after an enzymatic treatment with proteinase K and RNAse A. We conclude that the kit has a higher EV-derived RNA yield in comparison to our DU protocol, suggesting that it may be the method of choice for RNA sequencing purposes.