GSM8104683: milk EVs, control 4; Bos taurus; miRNA-Seq
The RNA was isolated from the EV using the miRNeasy Mini Kit (cat. No. 217004, QIAGEN, Germany). The isolation process followed the manufacturer's protocol. Briefly, EV samples were first lysed and homogenized in a highly denaturing guanidine-thiocyanate-containing buffer, which immediately inactivated RNases, ensuring the purification of intact RNA. By mixing with ethanol, the samples were loaded into an RNeasy Mini spin column. The total RNA bound to the membrane and contaminants were washed away. The eluted, high-quality RNA was collected and stored at -80 °C for further analysis. Briefly, libraries were subjected to reverse transcription and amplification by connecting 3' and 5' adapters to the ends of RNA molecules using T4 RNA Ligase (catalog no. LR2D1132K, Epicentre, US). cDNA constructs were purified and recovered, and the libraries were subsequently checked and normalized.