title: GSM4420259: H1944-LACZ-NoDOX_SMARCA2_R1; Homo sapiens; ChIP-Seq
library_construction_protocol: NCI-H1944 cells (20 million) stably transduced with doxycycline-inducible shSMARCA2 and either LACZ or SMARCA4 WT were treated with vehicle or 0.5 ug/mL doxycycline for 4 d to obtain significant SMARCA2 knockdown. Cells were fixed with 1% formaldehyde for 10 min, quenched with 0.125 M glycine for 10 min, washed with cold PBS three times and snap frozen. ChIP for SMARCA2 and SMARCA4 was performed by Active Motif Epigenetic Services. Chromatin was isolated with the addition of lysis buffer followed by disruption with a Dounce homogenizer. Lysates were sonicated, and DNA was sheared to an average length of 300-500 bp. ChIPs were performed with 30 ug of precleared chromatin and 5uL of anti-SMARCA2 (Abcam, ab15597) or 10uL anti-SMARCA4 antibody (Abcam, ab110641). Complexes were washed, eluted from the beads with SDS buffer and subjected to RNase and Proteinase K treatment. Crosslinks were reversed overnight at 65ºC, and DNA was purified by phenol-chloroform extraction and ethanol precipitation. Illumina-compatible libraries were generated using an automated system (Apollo 342, Wafergen Biosytems/Takara) and sequenced on the Illumina NextSeq 500 (single-end 75 bp reads).
attributes.tag: GEO Accession
attributes.value: GSM4420259