GSM6478616: TEAD4 ChIP liver YAP Tg biol rep 1; Mus musculus; ChIP-Seq
For ChIP-seq, ChIP was performed with Magna ChIP HiSens chromatin immunoprecipitation kit (Millipore Sigma) according to the manufacturer's protocols. Briefly, 100 mg mouse liver was pulverized with liquid nitrogen, and cross-linked in 1% methanol-free formaldehyde at room temperature for 10 minutes. Fixed cells were collected and lysed in Nuclei Isolation Buffer for 15 minutes. Then, cells were homogenized 10 times in a Dounce homogenizer (loose pestle). Extracted nuclei were fragmented by sonication to an average size of 200-700 bp (S2 Focused-ultrasonicator, Covaris). 10 μg fragmented chromatin was subjected to overnight incubation at 4 °C with specific antibody pre-conjugated beads. For ChIP normalization, 10 ng spike-in Drosophila chromatin (Active Motif, 53083) and 1 ug spike-in antibody (Active Motif, 61686) were added in each reaction before overnight incubation. After serial washing, the beads were incubated with ChIP elution buffer, Proteinase K at 65 °C for 2 hours and at 95°C for 15 minutes. The released DNA was purified using MinElute PCR Purification Kit (Qiagen). 5 ng of ChIP eluent were prepared using KAPA HTP Library Preparation Kit according to the manufacturer's protocols. PCR amplified libraries were purified with Ampure XP beads, then checked on the Agilent 2100 Bioanalyzer for quality control.