GSM3879730: Input_pooled_OPC; Homo sapiens; ChIP-Seq
Cells were fixed by adding 1/10 volume of freshly-prepared formaldehyde solution (11% formaldehyde, 0.1 M NaCl,  1 mM EDTA pH 8.0, 50 mM HEPES pH 7.9) to the existing media in each cell culture plate. Plates with added formaldehyde solution were agitated 15 minutes at room temperature. Fixation was stopped by adding 1/20 volume a 2.5 M glycine solution and incubating at room temperature for 5 minutes. Subsequently, cells were scraped off the culture dish surface and washed by transferring to a 50 ml conical tube on ice and pelleting at 800 x g in a refrigerated centrifuge for 10 minutes. Supernatant was removed and cells were re-suspended cells in 10 ml chilled PBS-Igepal (1x PBS pH 7.4, 0.5% Igepal CA-630). Cells were centrifuged again, supernatant was removed, and 10 ml chilled PBS-Igepal supplemented with 1 mM PMSF was added and the cell pellet was resuspemded. Cells were pelleted again at 800 x g, supernatant removed completely, and the cell pellet was snap-frozen on dry ice. Frozen cell pellets were kept at -80C and shipped to Active Motif (AM, commercial service provider) for further processing (chromatin extraction, fragmentation, antibody-precipitation and library preparation) according to AM's standard procedure. Libraries were prepared by Active Motif (Carlsbad, CA) for sequencing using standard Illumina protocols
GEO Accession
GSM3879730