GSM5229389: MDA-MB-231_Ctrl_1st; Homo sapiens; RNA-Seq
Total RNA was extracted with TRIZOL (Ambion). The RNA was further purified with two phenol-chloroform treatments and then treated with RQ1 Dnase (Promega, Madison, WI, USA) to remove DNA. And MCPIP1-RNA complex were isolated with MCPIP1 antibody. For each sample, 1μg of total RNA was used for RNA-seq library preparation. Polyadenylated mRNAs were purified and concentrated with oligo(dT)-conjugated magnetic beads (Invitrogen) before being used for directional RNA-seq library preparation. Purified mRNAs were fragmented at 95℃ in fragmentation buffer followed by end repair and 5' adaptor ligation. Then reverse transcription was performed with RT primer harboring 3' adaptor sequence and randomized hexamer. The cDNAs were purified and amplified and PCR products corresponding to 200-500 bps were purified, quantified