GSM8864922: Caco-2_Supernatant_S100_rep1; Homo sapiens; miRNA-Seq
For extracellular RNA isolation, media was collected and centrifuged to remove cell debris. Total exRNA was isolated from a portion of the supernatant by nucleic acid precipitation, followed by Trizol. The other portion of supernatant was centrifuged at 40,000xg. P40 exRNA was isolated from the resulting pellet using Trizol. The supernatant was collected and centrifuged at 100,000xg. S100 RNA was isolated from the supernatant by nucleic acid percipitation and subsequent Trizol isolation. P100 exRNA was isolated from the pellet using Trizol. Intracellular RNA was isolated from the Caco-2 cells using Trizol. sRNA libraries were constructed using the RealSeq-AC kit version 2 (Realseq Biosciences, Santa Cruz, CA, catalog no. 500-00048;) as per the manufacturer's instructions. We sent all sRNA libraries for sequencing to the University of Delaware Sequencing and Genotyping Center, where they were sequenced on a NovaSeq2000 instrument using 50-bp single-end reads.