exRNA reference profiling of 13 individuals addressing the effect of biofluid (serum vs. EDTA plasma), individual, gender, fasting and female hormonal cycle. exRNA reference profiling of 13 individuals addressing the effect of biofluid (serum vs. EDTA plasma), individual, gender, fasting and female hormonal cycle

Circulating extracellular RNAs (exRNAs) have the potential to serve as biomarkers for a wide range of medical conditions. However, limitations in existing exRNA isolation methods and a lack of knowledge on parameters affecting exRNA variability in human samples may hinder their successful discovery and clinical implementation. Using novel combinations of denaturants, reducing agents, proteolysis and revised organic extraction, we developed an automated, high-throughput approach for recovery of exRNAs and extracellular DNA (exDNA) from the same biofluid sample. We applied the method to characterize exRNAs from 312 plasma and serum samples collected from thirteen healthy volunteers at twelve time points over a two-month period. Overall design: sRNA-derived cDNA profiling of exRNA isolated from cell-free serum and EDTA plasma, using novel automated RNA isolation and multiplexed library preparation protocols. Biofluid samples were collected from 13 healthy individuals at 12 intervals over a period of two months.