GSM8395779: Tamoxifen, replicate 1; Homo sapiens; miRNA-Seq
Exosomes were isolated from serum-free media using Total Exosome Isolation Reagent kit (Invitrogen) based on the manufacturer's protocol with an increase in centrifuge time from 1 hour to 1.5 hours. Isolated exosomes were subsequently resuspended in 1 mL Takara Lysis Buffer and small RNAs were isolated using NucleoSpin miRNA kit (Takara Bio USA, Inc). The samples were further purified with RNeasy MinElute kit (Qiagen), with an increase in 100% ethanol volume from 250uL to 900uL. Sequencing libraries were prepared using the Illumina TruSeq Small RNA Library Prep Kit (Illumina) according to the manufacturer's protocol. A total of 6 sequencing libraries per run were pooled and normalized to 2 nM and denatured according to Illumina's NextSeq System Denature and Dilute Libraries Guide.