GSM6210181: Cardiomyocytes, Control, IgG, rep3; Mus musculus; OTHER
Adult cardiomyocytes were isolated from 10-12-week old mice by Langendorff perfusion. Cardiomyocyte fraction was enriched by low-speed centrifugation at 50g for 5 min.  H3K27ac Cut&Run was performed in cardiomyocytes as previously described . 200,000 cardiomyocytes are used for each Cut&Run experiment. H3K27ac (Cell Signaling Technology #8173) or Rabbit IgG (Cell Signaling Technology #3900) at 1:100 was incubated with cardiomyocytes which were attached to Concanavalin A coated magnetics beads (Bangs Laboratories #BP531) overnight at 4⸰C. 2.5 ul of pA-MNase from 1:10 dilution of the original stock provided by Henikoff lab was applied to cell-beads mixture and incubated at room temperature for 10min. Pull-down DNA was extracted in phase-lock tubes using phenol-chloroform method. DNA library was prepared using Hyper-prep kit (KAPA Biosystems KK8502) and submitted to Next-Generation Sequencing Core in University of Pennsylvania for sequencing. 40bp X 40bp paired-end sequencing was adopted.