title: GSM2175602: -LIF_2(RNA-Seq); Mus musculus; RNA-Seq
library_construction_protocol: RNA was extracted using TRIzol and purified on column with the RNeasy Mini Kit (Qiagen). Ribosomal RNA was depleted with the Ribo-Zero Gold rRNA Removal Kit (Illumina). RNA was lyophilized, suspended in 10 μL of water and fragmented to an average size of 200 base pairs by adding 5μL 1N NaOH (TEKnova) on ice for 30 minutes. The reaction was quenched with 25μL Tris HCl pH 7.0, brought up to 100 μL with water, and purified using the RNeasy Mini Kit (Qiagen). 3’ Phosphorylation, biotin-blocked adapter ligation, reverse transcription, immunoprecipitation, circularization, amplification, and PAGE separation were performed using the FAST-iCLIP library construction method as previously described (Flynn et. al 2014).
attributes.tag: GEO Accession
attributes.value: GSM2175602