title: GSM5654978: ChIP-seq_BMDM_LPS_BRG1_rep1; Mus musculus; ChIP-Seq
library_construction_protocol: ChIP-sequencing was performed as previously described  (doi: 10.1016/j.molcel.2012.10.013 ). In brief, 40 million cells were fixed with 2mM disuccinimidyl glutarate (DSG) at 4oC followed by 10 minutes fixation with 1% formaldehyde at RT. Cells were lysed in Fast IP buffer (150mM NaCl, 5mM EDTA, 5mM Tris, 1% Triton X-100 and 0.5% NP40) and cromatin was sheared with a Bioruptor in shearing buffer (1% SDS, 10µM EDTA and 0.05% Tris) until the majority of fragments were between 200-1000bp. Sheared chromatin was diluted 1:10 in Dilution buffer( 167mM NaCl, 16.7mM Tris, 1.2mM EDTA, 1.1% Triton X-100 and 0.01% SDS) and the IP was perfomed with 8µg of a-GR (Proteintech 24050-1-AP) and 16µg of a-BRG1(Cell Signalling #49360S and BRG1#abcam ab110641, 8µg from each) overnight. In the next day the IPs were incubated with Dynabeads (Invitrogen) for 6 hours. Afterwards the beads were washed with the FAST-IP budder and DNA was eluted in elution buffer (100mM NaHCO3 and 1%SDS). Library preperation was performed from 5 ng of ChIP DNA using the Kapa Hyper Prep Kit with PCR library amplification (#KK8504, KapaBiosystems) according to the manufactur's protocol. Shortly, ChIP DNA was end-repaired and A-tailed before adapter ligation. After adapter ligation, the ChIP DNA was purified with Agencourt AMPure XP beads (#A63880, Beckman Coulter) and size-selected for 360-600 bp using the Pippin Prep System from Saga Science. The size-selected library is amplified by PCR using the Kapa Hyper Prep Kit (#KK8504,KapaBiosystems) and purified terminally with Agencourt AMPure XP beads (#A63880, Beckman Coulter).