A protocol for rapid extraction of high-quality RNA from urinary extracellular vesicles

Efficient isolation of exosomes and other extracellular vesicles (EVs), and nucleic acids from urine presents challenges due to significant variability in the constituents of this biofluid, many of which are potent inhibitors of qRT-PCR. We present optimized workflows for isolation of both intact mRNA (and other long RNAs) and miRNA (and other short RNAs) from urinary EVs, and show their use for miRNA and mRNA biomarker detection. The novel workflow for isolation of exosomal RNA from urinary EVs avoids co-purification of inhibitors from samples and recovers RNA with high reproducibility.