GSM2704077: ChIP-SeqWT +IL15, anti-STAT5B; Mus musculus; ChIP-Seq
For ChIP-Seq, and cross-linkedWith 1% formaldehyde (methanol-free, Pierce, Rockford, IL) at room temperature for 10 min. After sonication, fragmented chromatin equivalent to 10 million cellsWas immunoprecipitatedWith control rabbit IgG (Santa Cruz Biotechnology, Dallas TX) or anti-STAT5B (R&D systems) and Magna ChIPTM Protein A+G Magnetic Beads (Millipore, Billerica MA). ChIP-Seq DNA librariesWere prepared using KAPA LTP Library Preparation Kit (Kapa Biosystems,Wilmington, MA) and indexed primers (BIOO Scientific, Austin, TX), and the librariesWere then sequenced. For RNA-Seq, KAPA Stranded RNA-Seq Library Preparation Kit (KAPA Biosystems,Wilmington, MA), and each libraryWas indexed using barcoded primers (BIOO Scientific, Austin, TX). Barcoded PCR productsWere purified on 2% E-Gel, 250-400 bp fragmentsWere purified, quantified on Qbit (Invitrogen), mixed, and sequenced on an Illumina HiSeq2000 platform. PCR productsWere barcoded (indexed) and sequenced on Illumina HiSeq 2000 platform.