Urinary extracellular vesicles for RNA extraction: optimization of a protocol devoid of prokaryote contamination.

ABSTRACT:

Background

Urinary extracellular vesicles (UEVs) represent an ideal platform for biomarker discovery. They carry different types of RNA species, and reported profile discrepancies related to the presence/absence of 18s and 28s rRNA remain controversial. Moreover, sufficient urinary RNA yields and respective quality RNA profiles are still to be fully established.

Methods

UEVs were enriched by hydrostatic filtration dialysis, and RNA content was extracted using 7 different commercially available techniques. RNA quantity was assessed using spectrophotometry and fluorometry, whilst RNA quality was determined by capillary electrophoresis.

Results

The presence of prokaryotic transcriptome was stressed when cellular RNA, as a control, was spiked into the UEVs samples before

SUBMITTER: Tataruch-Weinert D 

PROVIDER: S-EPMC4921785 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature