title: GSM9458944: TEAD1 ChIPseq; Homo sapiens; ChIP-Seq
library_construction_protocol: 1,000,000 HCASMCs were cross linked in 1% formaldehyde for 10 minutes and washed with PBS and replaced with hypotonic buffer (20 mM Hepes (pH 7.9), 10 mM KCl, 1 mM EDTA (pH 8) and 10% glycerol) and incubated on ice for 6 minutes. Cells were then sonicated using a Branson 250 Sonifier (using power setting 5, constant duty for 10 rounds of 30-second pulses) with confirmation of chromatin fragments at 250-400 babse pairs. Lysates were then incubated overnight with 5 µg of anti-CEBPB (Santa Cruz sc-150), TEAD(Cell Signaling 12292S), or H3K27ac(Abcam 4729). Protein-DNA complexes were captured with Protein G agarose beads (Sigma 8104) and eluted in 1% SDS TE buffer at 65 °C. After reverse cross-linking followed by RNase A and proteinase K digestion, chromatin was purified using a QIAquick PCR purification kit (Qiagen 28706). ChIP DNA sequencing libraries were prepared using the Kappa HyperPrep (Roche 07962347001)