Development and application of a scalable workflow for immunomagnetic separation of exRNA carrier subclasses and molecular analysis of their cargo.
Our aim with this study was to evaluate the performance of immunomagnetic separation, differential ultracentrifugation and size exclusion chromatography for the isolation of different extracellular RNA carriers from biofluids. Serum and plasma were collected from healthy donors and pooled. Placental explants from healthy pregnancies were cultured and supernatant was collected. Extracellular RNA carriers were isolated using various methods including immunomagnetic separation, differential ultracentrifugation and size exclusion chromatography. Isolated RNA was subjected to small RNA library preparation and analyzed using small RNA seq to compare the efficiency and reproducibility of the different extracellular RNA carrier separation methods. Raw fastq files from the sequencing experiments... (for more see dbGaP study page.)