title: GSM2671193: KO-Brg1 ChIP; Mus musculus; ChIP-Seq
library_construction_protocol: 50 ug total fragmented chromatin were mixed with 5 ug antibody overnight at 4 degree with rotation. The chromatin-antibody complex were then precipitated with protein A/G magnetic beads (Thermo Fisher Scientific). After washes with low salt wash buffer (20mM Tris, 2mM EDTA, 50mM NaCl, 0.01% SDS and 1% SDS), high salt wash buffer (same composition, 250 mM NaCl) and LiCL-wash buffer (10mM Tris, 1mM EDTA, 250 mM LiCl, 1% NP40 and 1% sodium deoxycholate), precipitated chromatin was eluted in 200µl of elution buffer (10µl 20% SDS, 20µl 1M NaHCO3 and 170µl dH2O). Reverse cross linking was done by adding 8 ul 5M NaCl and incubate at 65 deg overnight. 1 ul 10 mg/ml RNase A was added to the tube for 30 mins incubation at 37°C. Then 4µl 0.5M EDTA, 8µl 1M Tris-HCl and 1µl 20 mg/ml Proteinase K was added for 2h incubation at 42°C. The released DNA was purified using QIAquick PCR purification kit (Qiagen). Purified DNA from two replicates were pooled for library preparation using TruSeq Nano DNA Library Prep Kit (Illumina) according to the manufacturer's instructions.
attributes.tag: GEO Accession
attributes.value: GSM2671193