GSM5949422: EVs from HUVEC LNP-treated rep1; Homo sapiens; RNA-Seq
EVs were isolated from cell culture medium by ultracentrifugation. To removed cell debris, the cell culture medium were centrifuged at 3000 x g for 15 min at 4°C, ultracentrifuged at 60,000 x g for 35 min at 4°C, followed by filtration through 0.2μm filters at 60,000 x g for 30 minutes at 4 °C. The EV pellet were collected and resuspended in PBS after a second round of ultracentrifugation at 120,000 x g for 70 minutes at 4°C. Library construction was performed using Takara SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian kit, which is specifically designed for very low input total RNA samples.