GSM2221657: U2AF2_control iCLIP 1; Homo sapiens; RIP-Seq
For iCLIP protocol, nuclear extracts (detergent lysis) were prepared from  cells (control or T7-hnRNPA1 inducible)  prior to immunoprecipitation.  For RNA-seq, whole cell extracts (detergent lysis) were prepared from cells (control or T7-hnRNPA1 inducible)  and RNA was isolated using TRIreagentLS. For iCLIP, libraries were constructed as previously described (Huppertz, I., et al. Methods. 2014) with minor changes. 3' Adenylated adapter, AIR ligase, and PCR primers used from NEXTflex™  Illumina Small RNA-Seq Library Prep Kit (Bioo Scientific) and RT primers were adapted for kit reagent use. For RNA-seq, and Panc1 RIP-seq,  NEXTflex™ Rapid Directional qRNA-Seq™ Kit was used (Bioo Scientific). For small RNA-seq library prepartation, NEXTflex™ Small RNA-Seq Kit v1 was used. For iCLIP, RNA purification was performed as previously described (Huppertz, I., et al. Methods. 2014). For RNA-seq, PolyA+ selection was performed.
GEO Accession
GSM2221657