Rigor and Reproducibility of Acoustofluidics of saliva from healthy donors

This is for Rigor and Reproducibility study to demonstrate the reproducibility of AFS SOP for EV separation, RNA isolation, and RNA analysis by using saliva samples from healthy individuals. Blinded saliva samples have been processed by two independent AFS operators for EV separation and subjected to miRNA-seq analysis. EV-associated miRNA seqeunces were used as a monitoring tool for Rigor and Reproducibility of AFS process between operators. Saliva samples from healthy volunteers (5 female and 5 male) were collected based on our research SOP and split into two fractions for AFS analysis by two independent operators. Resulting two AFS fractions (waste and collected) per sample per operator were blinded and sent to UCSD for miRNA-seq analysis. Resulting sequencing data have been analyzed for the statistical evaluation of Rigor and Reproducibility in AFS separation of EV’s. This has been determined by evaluating the statistical difference in miRNA’s existing in EV’s separated from each sample by two independent operators.