GSM2196891: UACC-62_Input_R1; Homo sapiens; RIP-Seq
RNA was extracted after immunoprecipitation using TRI Reagent (Sigma) including DNase I treatment step. An estimated mass of 20 ng of RNA per sample was used. Ribosomal RNAs were removed with the Ribo-Zero Gold Kit (Epicentre Cat. No. RZHM11106/RZG1224) and randomly fragmented, converted to double stranded cDNA and processed through subsequent enzymatic treatments of end-repair, dA-tailing, and ligation to adapters as in Illumina's "TruSeq RNA Sample Preparation v2 Protocol" (Part # 15026494 Rev. C). Adapter-ligated library was completed by 10 cycles of PCR with Illumina PE primers. The resulting purified cDNA library was applied to an Illumina flow cell for cluster generation (TruSeq cluster generation kit v5) and sequenced on the Genome Analyzer IIx with SBS TruSeq v5 reagents by following manufacturer's protocols.