title: GSM2044513: ChIP-seq of V6 5 SHC Input in mESC; Mus musculus; ChIP-Seq
library_construction_protocol: For ChIP-Seq, 5×107 cells were used per immunoprecipitation according to previously described protocols (Lee et al., 2006). ChIP-Seq libraries were prepared with Illumina's ChIP-Seq sample prep kit (0801-0303).ES cells (V6.5) were infected with lentivirus carrying either Non-targeting shRNA or ZFP281 shRNA in the presence of 8 ug/ml of polybrene. 24 hours later, ES cells were selected with 2 ug/ml of puromycin for additional 48 hours and then were grown one passage off feeders for 30 min before harvesting. Total RNA was isolated with the RNeasy (74106 Qiagen) kit, treated with DNase I (M0303L NEB), and re-purified with RNeasy. For total RNA-seq analyses, 2.5 µg of total RNA was depleted of ribosomal RNA with the Ribo-Zero kit from Epicentre (#RZH1046). Ribosomal RNA-depleted samples were used for library preparation with the TruSeq RNA Sample Prep Kit (Illumina) for next-generation sequencing. NEBNext® DNA Library Prep Master Mix Set for Illumina; TruSeq RNA Sample Prep Kit (Illumina)
attributes.tag: GEO Accession
attributes.value: GSM2044513