Whole Transcriptome Analysis (Rna-Seq) By Next-Generation Sequencing of hAF-EVs
II trimester human amniotic fluid (hAF) samples were collected as leftover material from prenatal screening. hAF samples were concentrated and processed by size exclusion chromatography (SEC). hAF-EV-enriched fractions were collected and concentrated by ultracentrifugation and then resuspended in a PBS solution. Combined phenol/guanidine-based lysis of samples and silicamembraneâ€“based purification of total RNA. (miRNeasy Micro Kit, Qiagen)  Whole transcriptome RNA-seq analyses have been performed using the Universal RNAseq kit (Nugen, Tecan).