GSM7622293: Ma_1064,Female,ΔEcad,H3K27ac; Mus musculus; ChIP-Seq
Cells were fixed in 10mL 0.4% PFA for 10 minutes at RT. Crosslinking reaction was stopped by addition of 1 mL 2M Glycine (pH= 8.0). Cells were resuspended in 1 mL PBS and sonicated until DNA being fragmented to an average of 400 bp. Chromatin was harvested using an anti-H3K27ac (Active motif #39133) and protein G sepharose beads. DNA was purified by protein digestion by Protein K and phenol/chloroform (1:1) extraction. Libraries were prepared using the Illumina TruSeq ChIP library preparation kit following the manufacturer's protocol. 2 to 3 ng of DNA were subjected to subsequent steps of end-repair, dA-tailing and ligation of TruSeq RNA UD indexes Illumina adapters. Librairies were amplified using 15 cycles on the thermocycler.