title: GSM8767857: SCAC-Pooled_Input; Homo sapiens; ChIP-Seq
library_name: GSM8767857
library_construction_protocol: Cells were fixed for 15 min in 1/10 volume Formaldehyde solution (11% Formaldehyde, 0.1M NaCl, 1mM EDTA pH 8.0, 50mM Hepes pH 7.9 in milliQ H2O) at room temperature. Fixation was stopped by adding 1/20 volume glycine solution (2.5M in milliQ H2O) for 5 min at room temperature. Cells were then scraped, transferred into 50mL conical tubes, and kept on ice. Tubes were centrifuged at 800xg at 4°C for 10 min. Pelleted cells were then resuspended in 10mL of chilled PBS-Igepal solution (0.5% Igepal in 1X PBS). Cells were repelleted as before and washed twice more with PBS-Igepal, adding 100uL of PMSF (1mM in ethanol) after the second wash to each sample. Pellets were then snap frozen on dry ice.Further processing for ChIP sequencing and analysis, including chromatin extraction, fragmentation, antibody precipitation, and library preparation was performed commercially by Active Motif Services using a validated antibody to BATF2 (Proteintech 16592-1-AP). Sequence libraries were generated using the standardized steps of enzymatic end polishing, dA-addition, and adaptor ligation. Following PCR amplification, the resulting libraries were quantified, sequenced, and quality filtered using Illumina software (75 nucleotide reads, single end).
identifiers.namespace: GEO
identifiers.id: GSM8767857_r1