title: GSM7830905: treated with pre-miR203a vector rep2.RNA; Homo sapiens; RNA-Seq
library_construction_protocol: MCF10A cells stably expressing pMSCVpuro-pre-miR-203a or pMSCVpuro were lysed for 30 minutes on ice in cell lysis buffer then centrifuged for 30 minutes at 14,000 rpm at 4°C. The supernatant was used as cell lysate for the IP. Mouse anti-AGO2 (Abcam) was coupled to Protein G Dynabeads (Thermo Fisher Scientific) and added to lysates. Antibody-lysate mixtures were rotated for 4 hours at 4°C [to bind RISC in each sample]. The beads were washed in IP wash buffer supplemented with a protease inhibitor cocktail and eluted in Trizol (Invitrogen). mRNA purification, cDNA synthesis, and amplification were carried out according to the TrueSeq RNA sample preparation guide (Illumina).