Mouse Embryonic Stem Cells E14 ChIP-seq (union of Day 0, Day 15 and two Day 30 samples)
Cells in different time points of the adipocyte differentiation from mES E14 were cross-linked in two steps using first Disuccinimidyl glutarate (DSG) for 30 minutes then 1 % methanol-free ultrapure formaldehyde for 10 minutes at room temperature. Glycine was added for 5 minutes in 125 mM final concentration. After fixation chromatin were sonicated to generate 200-1000 bp fragments. Chromatin was immunoprecipitated with antibodies against H3K27ac (Abcam, ab4729). The eluted DNA was purified and quantified with Qubit fluorometer. ChIP-seq libraries were prepared by Ovation Ultralow Library Systems (Nugen) according to manufacturer's instructions. Briefly, 1ng immunoprecipitated DNA were submitted to End-repair, then adaptors were ligated to double stranded DNA fragments. Adaptor containing fragments were PCR amplified in 18 cycles. Libraries were gel extracted and analyzed by Agilent 1000 DNA chip. Libraries were sequenced on HiScanSQ according to manufacturer's protocol. This is a merge of the four samples (Day 0, Day 15, Day 30 Rep1, Day 30 Rep2).