NextSeq 500 sequencing; Small RNA-sequencing of plasma exosome-enriched extracellular vesicles derived miRNAs from lactating mothers with type 1 diabetes and lactating healthy mothers
Small RNA-sequencing of plasma exosome-enriched extracellular vesicles derived miRNAs from lactating mothers with type 1 diabetes and lactating healthy mothers
Briefly, 3 ml plasma per sample was diluted with 3 ml of 1X PBS (pH 7.5) followed by centrifugation at 5,000 x g for 3 min. Next, 3.5 ml of the supernatant was filtered with a nitrocellulose filter (0.8 µm pore size, VWR, Radnor, PA, USA). The extracellular vesicles were extracted by ExoEasy Serum Plasma Kit (Qiagen, Valencia, CA, USA) according to the manufacturer's protocol.  The exosomal RNA were extracted by exoRNeasy Serum Plasma Kit (Qiagen) Libraries were prepared using QIAseq miRNAseq library kit following the manufacturer's instructions.